CN107515187A - A kind of method of vessel cell morphological feature in quick detection lignocellulosic material - Google Patents
A kind of method of vessel cell morphological feature in quick detection lignocellulosic material Download PDFInfo
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- 239000012978 lignocellulosic material Substances 0.000 title claims abstract description 74
- 230000000877 morphologic effect Effects 0.000 title claims abstract description 74
- 238000001514 detection method Methods 0.000 title claims abstract description 63
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- 238000012545 processing Methods 0.000 claims abstract description 46
- 238000002360 preparation method Methods 0.000 claims abstract description 10
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- 239000000463 material Substances 0.000 claims description 41
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 9
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 claims description 8
- 239000002771 cell marker Substances 0.000 claims description 8
- 230000018044 dehydration Effects 0.000 claims description 8
- 238000006297 dehydration reaction Methods 0.000 claims description 8
- 239000000975 dye Substances 0.000 claims description 7
- 238000006243 chemical reaction Methods 0.000 claims description 6
- 229920006395 saturated elastomer Polymers 0.000 claims description 6
- 238000004043 dyeing Methods 0.000 claims description 5
- 230000003287 optical effect Effects 0.000 claims description 5
- 230000009286 beneficial effect Effects 0.000 claims description 4
- 239000003124 biologic agent Substances 0.000 claims description 3
- 239000003086 colorant Substances 0.000 claims description 3
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- 239000002657 fibrous material Substances 0.000 claims description 3
- 238000007689 inspection Methods 0.000 claims description 2
- 238000010998 test method Methods 0.000 abstract description 2
- 241000219000 Populus Species 0.000 description 18
- 235000017166 Bambusa arundinacea Nutrition 0.000 description 15
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Abstract
The present invention relates to a kind of method of vessel cell morphological feature in quick detection lignocellulosic material, belong to lignocellulosic material microstructure detection technique field.This method is to obtain micro-image first, and sample preparation early stage is carried out to lignocellulosic material；The sample prepared is obtained to the micro-image that can be told each lignocellulosic material and respectively form cell by light microscope；Image pixel is carried out to the micro-image of acquisition and actual size proportionate relationship determines and processing, to ensure the readability and the accuracy of identification and detection of vessel cell morphological feature in micro-image；The parameter of micro-image vessel cell morphological feature detection after setting processing；Vessel cell morphological feature detection is carried out, it is final to obtain vessel cell image and vessel cell characteristic pattern data in the lignocellulosic material for meeting setup parameter requirement.The present invention, which has filled up current method of testing, can not obtain the morphological feature such as lignocellulosic material vessel cell number, area and circularity this blank automatically.
Description
Technical field
It is wooden especially with regard to a kind of quick detection the invention belongs to lignocellulosic material microstructure detection technique field
The method that fibrous material forms cellular vessel cell morphological characteristic.This method can utilize the fast automatic acquisition of image analysis method
The morphological feature parameter of vessel cell in lignocellulosic material micro-image.
Background technology
Lignocellulosic material is reproducible green building and woodwork raw material, while is also biomass energy material
Important raw and processed materials.Lignocellulosic material mainly includes timber, bamboo wood and windmill palm vine material.The wood fibre material during utilization
It is the xylem part within bark to expect what is mainly utilized.Lignocellulosic material xylem is by fiber or tracheid, vessel cell, thin
The polytype such as parietal cell and wood radiaftive rays cell forms, and wherein fiber or tracheid mainly plays mechanical support in xylem,
And in addition to softwood tracheid has the function of vessel cell, vessel cell is respectively provided with other lignocellulosic material xylems,
Vessel cell has important transporting moisture function, and dissection important in forest microtexture in the growth course of forest
The factor.
Lignocellulosic material xylem is because by various types of cell arrangement, it not only forms a variety of
The grain of wood, and its different microstructure and its dissection property also determine the physical mechanics property of lignocellulosic material
Energy, finally determine the purposes of lignocellulosic material.It can be seen that different lignocellulosic material micro-structural features be identification timber with
And understand the basic of its material characteristic, therefore lignocellulosic material micro-structural feature this respect research since microscope send out
Obtain constantly carrying out always since bright.Vessel cell is the dissection factor important in lignocellulosic material microstructure, accurate
Really the morphological feature of detection vessel cell and its shared proportion in microstructure are particularly important.
The morphological feature measuring method on lignocellulosic material fiber and vessel cell can be divided into two classes, Yi Leifang at present
Method is chemistry isolation process or physics stripping method, and the method is by the composition cell sepn of xylem or is stripped out, and it is complete to select form
Whole cell carries out the measurement of its longitudinal length and transverse width, and measurement emphasis is mainly fiber properties, especially suitable for pulpwood
The evaluation of fiber properties；Another kind of method is in site measurement method, is that current in site measurement lignocellulosic material fiber and conduit are thin
The main method of the morphological feature such as the wall thickness of born of the same parents and chamber footpath.Material is made into section first in site measurement method or acquisition surface is clear
Structure, which is directly taken pictures, obtains micro-image, then carries image analysis software using microscope under the microscope and uses linear survey
Method, will be located at vessel cell center homonymy inner surface contour line on a little and outer wall contour line on a little between straight line away from
From as vessel cell wall thickness, using the air line distance of the point-to-point transmission on the inner surface contour line of vessel cell center both sides as leading
Solencyte chamber footpath.The method can only measure fiber and vessel cell cell one by one, and due to the form of vessel cell
It is not the circle of standard, vessel cell area value and entirely micro- can not also be obtained after the value of vessel cell chamber footpath exactly by obtaining
Area proportion in image shared by vessel cell.
Image J are a kind of powerful can to freely download the image analysis program used.Led at present in biology
Domain is widely used to the area estimation of cell count, quantitative analysis fluorescence intensity, vessels analysis and single irregularly shaped object
Etc..Vessel cell due to being surrounded by all types of cells such as fibrocyte and parenchyma cell, different from it is common it is single from
Scattered cell, therefore can not accurately obtain the micro- structure of lignocellulosic material the methods of existing cell count using Image J
Make middle vessel cell morphological feature parameter.
The content of the invention
It is a primary object of the present invention to overcome the weak point of prior art, there is provided a kind of quick complete detection is wooden
The method of vessel cell morphological feature in fibrous material.Such a method quickly can be extracted defeated in lignocellulosic material composition comprehensively
The image of histocyte conduit morphological feature is led, while its number, area, circularity and in lignocellulosic material group can be obtained
The morphological features such as the area ratio into cell shared by it.
The present invention adopts the following technical scheme that：
A kind of method of vessel cell morphological feature in quick detection lignocellulosic material, it is characterised in that this method is first
Before this to obtain micro-image, sample preparation early stage is carried out to lignocellulosic material；The sample prepared is passed through into optical microphotograph
Mirror, which obtains, can tell the micro-image that each lignocellulosic material respectively forms cell；Image pixel is carried out to the micro-image of acquisition
With actual size proportionate relationship determine and handle, for ensure micro-image in vessel cell morphological feature readability and identification and
The accuracy of detection；The parameter of micro-image vessel cell morphological feature detection after setting processing；Carry out vessel cell form
Feature detection, it is final to obtain vessel cell image and vessel cell form in the lignocellulosic material for meeting setup parameter requirement
Characteristic.
This method specifically includes following steps：
1) to obtain micro-image, sample preparation early stage is carried out to lignocellulosic material：
According to lignocellulosic material species and shape, it is cut into beneficial to the block-shape morphology material for obtaining micro-image, the block
Shape material obtains as sample for its micro-image；
2) each lignocellulosic material sample can be told by, which obtaining the sample prepared by light microscope, respectively forms carefully
The micro-image of born of the same parents：
The sample that step 1) is prepared cuts the section that thickness is 10-30 μm；Then with biological agent dyestuff to cut into slices into
Row dyeing, to guarantee to distinguish the different types of each histocyte for forming lignocellulosic material, dyeing time is according in light
Depending on each histiocytic clear-cut required duration observed under microscope；Then cut into slices by graded ethanol
In each tissue cell dehydration, and each histocyte after dehydration is changed into transparent using dimethylbenzene, with mountant to each tissue
Section after cell is changed into transparent carries out mounting processing；Section after mounting processing is observed under an optical microscope takes pictures, and obtains
The micro-image of lignocellulosic material sample, for all types of lignocellulosic material samples, this can be distinguished by preserving each sample
The micro-image of vessel cell in sample；
3) image pixel is carried out to the micro-image of acquisition and actual size proportionate relationship determines and image procossing, to ensure
The readability and the accuracy of identification and detection of vessel cell morphological feature, are specifically included in micro-image：
3.1) image pixel and actual size dependency relation ratio are determined：The ratio on micro-image that will be obtained through step 2)
The physical length of example chi is corresponding with the pixel of the engineer's scale, you can the proportionate relationship between image pixel and actual size is determined,
For ensuring that what is obtained when detecting vessel cell morphological feature is the actual size of conduit；
3.2) contrast enhancement processing：By adjusting image pixel and actual size dependency relation ratio are determined through step 3.1)
The image saturated pixel value of the micro-image of the lignocellulosic material sample of example, strengthens vessel cell and fiber in the micro-image
The contrast of cell and other cell types, contribute to the identification of vessel cell morphological feature；
3.3) image type conversion process：Image after step 3.2) contrast enhancement processing is converted into binary system ash
Spend image, the i.e. only image of two kinds of hue distinguishes of black and white；
3.4) ductal cell marker is handled：The binary system gray level image that step 3.3) obtains is entered by selected threshold to two
Vessel cell is marked in gray level image processed, and respectively tissue is thin for the lignocellulosic material in the binary system gray level image of different threshold values
Color that born of the same parents show is different, is set as optimal threshold when all designated colors of the only chamber of vessel cell and wall color
Vessel cell in the image has been carried out mark processing by value, the micro-image obtained by the optimal threshold, this mark
It is easy to the detection subsequent to vessel cell；
4) the micro-image vessel cell morphological feature detection parameters after setting processing, are specifically included：
4.1) vessel cell area parameters are set：Select through step 3) processing after micro-image in larger vessel cell and
Compared with ductule cell, its diameter is measured respectively, and calculates the area S ' of larger vessel cell in fibre imagemaxWith compared with ductule
The area S ' of cellmin；The vessel cell area lower limit S of settingmin＜ S 'min, the vessel cell area higher limit S of settingmax
＞ S 'max, and the two round numbers；The vessel cell area S to be detected then set meets：Smin≤S≤Smax；
4.2) vessel cell circularity parameter setting：It is thin according to conduit to be detected in the micro-image after step 3) processing
The circularity setting vessel cell circularity parameter c to be checked measured of born of the same parents, 0 ＜ c≤1, wherein, 1 represents the circle of rule；
5) vessel cell morphological feature detects：After vessel cell morphological feature detection parameters according to step 4) setting, obtain
The vessel cell image and vessel cell characteristic pattern data of detection parameters requirement must be met；The vessel cell morphological feature
Data include：Vessel cell morphological data overall information, single vessel cell numbering and corresponding size and circularity form
Information, it is finally completed the detection of lignocellulosic material vessel cell morphological feature.
The features of the present invention and beneficial effect：
The invention provides a kind of method of vessel cell morphological feature in quick detection lignocellulosic material, such a method
The image of conducting tissue cellular vessel cellular morphology in being formed with rapid extraction lignocellulosic material, while can quickly, certainly
Move and accurately analyze individual or multiple lignocellulosic material displaing micro picture vessel cell shape informations, obtain its number, area, circle
The area ratio of morphological feature and the vessel cells such as degree in wood materials form cell shared by it.Fill up current traditional
Method of testing can not obtain the morphological feature such as lignocellulosic material vessel cell number, area and circularity this blank automatically.This
Inventive method carries out rapid, high volume screening for genetically modified tree and directive breeding forest has important practical application meaning.
Brief description of the drawings
Fig. 1 is the FB(flow block) of the present invention.
Fig. 2 is the binary system gray-scale map of three kinds of lignocellulosic material cross sections provided in an embodiment of the present invention microstructure；
Fig. 2 (a) is poplar, and Fig. 2 (b) is Mao bamboo timber, and Fig. 2 (c) is herba fibraureae recisae material.
Fig. 3 is the ductal cell marker figure with three kinds of lignocellulosic material microstructures corresponding to Fig. 2.
Fig. 4 be with Fig. 3 corresponding to vessel cell Detection and Extraction figure in three kinds of lignocellulosic materials.
Embodiment
It is further described below by specific embodiment and accompanying drawing in the quick detection lignocellulosic material of the present invention
Possessed technique effect in the content and feature and application of vessel cell morphological feature method, but the present invention not therefore and by
To any restrictions.
The invention provides a kind of method of vessel cell morphological feature in quick detection lignocellulosic material, this method are first
Before this to obtain micro-image, sample preparation early stage is carried out to lignocellulosic material；The sample prepared is passed through into optical microphotograph
Mirror, which obtains, can tell the micro-image that each lignocellulosic material respectively forms cell；Image pixel is carried out to the micro-image of acquisition
With actual size proportionate relationship determine and handle, for ensure micro-image in vessel cell morphological feature readability and identification and
The accuracy of detection；The parameter of micro-image vessel cell morphological feature detection after setting processing；Carry out vessel cell form
Feature detection, it is final to obtain vessel cell image and vessel cell form in the lignocellulosic material for meeting setup parameter requirement
Characteristic.
The flow chart of the inventive method specifically includes following steps referring to Fig. 1：
1) to obtain micro-image, sample preparation early stage is carried out to lignocellulosic material：
According to lignocellulosic material (generally including timber, bamboo wood and rattan material) species and shape, it is cut into beneficial to acquisition
Lignocellulosic material (such as is cut into cross-sectional area and is less than or equal to 10mm by the block-shape morphology material of micro-image2, longitudinal direction it is long
Spend the cylindrical material for 15mm), the bulk material obtains as sample for its micro-image.
2) each lignocellulosic material sample can be told by, which obtaining the sample prepared by light microscope, respectively forms carefully
The micro-image of born of the same parents：
The sample that step 1) is prepared cuts the section that thickness is 10-30 μm；Then with biological agent dyestuff (for example with
Sarranine, fast green or toluidine blue) section is dyed, form the different types of of lignocellulosic material to guarantee to distinguish
Each histocyte, dyeing time according under an optical microscope observe each histiocytic clear-cut required duration and
It is fixed；Then each tissue cell dehydration in being cut into slices by graded ethanol, and make each tissue after dehydration thin using dimethylbenzene
Born of the same parents are changed into transparent, and the section after being changed into transparent to each histocyte with mountant carries out mounting processing；Section after mounting processing
Observation is taken pictures under an optical microscope, the micro-image of lignocellulosic material sample is obtained, for all types of lignocellulosic materials
Sample, the micro-image of vessel cell in the sample can be distinguished by preserving each sample.But the present invention obtains the side of micro-image
Method is not therefore any way limited.
3) image pixel is carried out to the micro-image of acquisition and actual size proportionate relationship determines and image procossing, to ensure
The readability and the accuracy of identification and detection of vessel cell morphological feature, are specifically included in micro-image：
3.1) image pixel and actual size dependency relation ratio are determined：The ratio on micro-image that will be obtained through step 2)
The physical length of example chi is corresponding with the pixel of the engineer's scale, you can the proportionate relationship between image pixel and actual size is determined,
It is the actual size of conduit (such as by IMAGE J images point for ensure to obtain when detecting vessel cell morphological feature
Engineer's scale physical length on micro-image is inputted when analysing the setting of program medium scale, while long measure is set as " μm ", is examined
The vessel cell morphological data obtained during survey will be with " μm " for unit)；
3.2) contrast enhancement processing：By adjusting image pixel and actual size dependency relation ratio are determined through step 3.1)
The image saturated pixel value of the micro-image of the lignocellulosic material sample of example, strengthens vessel cell and fiber in the micro-image
The contrast of cell and other cell types, contribute to the identification of vessel cell morphological feature (such as by IMAGE J images point
Analyse program and carry out contrast enhancement processing, the image saturated pixel value scope of selection is 0.2-0.8, and it is thin to obtain conduit after processing
Born of the same parents and other cell types contrast obvious image)；
3.3) image type conversion process：Image after step 3.2) contrast enhancement processing is converted into binary system ash
Image is spent, i.e., only (it is 8- such as to select image type by IMAGE J image analysis programs to the image of two kinds of hue distinguishes of black and white
Bit types)；
3.4) ductal cell marker is handled：The binary system gray level image that step 3.3) obtains is entered by selected threshold to two
Vessel cell is marked in gray level image processed, and respectively tissue is thin for the lignocellulosic material in the binary system gray level image of different threshold values
Color that born of the same parents show is different, is set as optimal threshold when all designated colors of the only chamber of vessel cell and wall color
Vessel cell in the image has been carried out mark processing by value, the micro-image obtained by the optimal threshold, this mark
Being easy to the detection subsequent to vessel cell, (such as by IMAGE J image analysis programs, gray level image optimal threshold is in 50-255
Chosen in scope, during optimal threshold, the chamber and all red of wall color of vessel cell)；
4) the micro-image vessel cell morphological feature detection parameters after setting processing, are specifically included：
Need to set to be detected lead before carrying out the micro-image conduit morphological feature detection of lignocellulosic material sample cross section
Solencyte morphological feature parameter, is divided into conduit area parameter setting and circularity parameter setting：
4.1) vessel cell area parameters are set：Select through step 3) processing after micro-image in larger vessel cell and
Its diameter d is measured compared with ductule cell and respectively (due to image pixel and actual size dependency relation ratio, it is known that the center of circle excessively exists
Vessel cell both sides inwall draws a straight line, you can obtains its diameter actual size).It is assumed that circle of the vessel cell for rule, according to
According to formula π × (d/2)2Substantially calculate larger vessel cell area S ' in micro-imagemaxWith compared with ductule cell area S 'min,
Primarily determine that vessel cell areal extent numerical value [S 'min,S’max], in order to not omit minimum vessel cell and maximum vessel cell,
The vessel cell area lower limit S being normally set upmin＜ S 'min, the vessel cell area higher limit S of settingmax＞ S 'max, generally
The two round numbers；The vessel cell area S to be detected then set meets：Smin≤S≤Smax, and S is usually integer；
4.2) vessel cell circularity parameter setting：It is thin according to conduit to be detected in the micro-image after step 3) processing
Circularity (circularity) the setting vessel cell circularity parameter c to be checked measured of born of the same parents, 0 ＜ c≤1, wherein, 1 represents rule
It is circular (as utilized IMAGE J image analysis programs, to set suitable vessel cell circularity scope in circularity scope between 0-1
With c, 0 ＜ c≤1, such as selected 1, it is standard circular that can detect vessel cell；4 π of IMAGE J image analysis programs foundation ×
[vessel cell area]/[vessel cell girth]2Formula determines each vessel cell circularities)；
5) vessel cell morphological feature detects：After vessel cell morphological feature detection parameters according to step 4) setting, obtain
The vessel cell image and vessel cell characteristic pattern data of detection parameters requirement must be met；The vessel cell morphological feature
Data include：Vessel cell morphological data overall information (including detect vessel cell total quantity, the gross area, average area,
The area ratio of average roundness and all vessel cells detected shared by whole micro-image), single vessel cell
Numbering and corresponding size and circularity shape information, the detection of lignocellulosic material vessel cell morphological feature has been finally completed it
The detection parameters of (such as according to step 4.1) and 4.2) input obtain detection knot by performing IMAGE J graphical analysis particles program
Fruit).
Embodiment 1
1) to obtain poplar micro-image, sample preparation early stage is carried out to poplar material：
According to lignocellulosic material broadleaf poplar shape, cross-sectional area is less than 10mm2Poplar be cut into longitudinal length
For the cylindrical material of 15mm length, the bulk material obtains as sample for its micro-image.
2) sample prepared is obtained to the micrograph that can be told poplar sample and respectively form cell by light microscope
Picture：
The sample that step 1) is prepared, timber ordinary strip method is used to cut thickness as 20 μm of section；Then with 1%
Sarranine aqueous dyestuff dyes to section, and dyeing time is 10-12 hours；Then in being cut into slices by graded ethanol
Each tissue cell dehydration, and each histocyte after dehydration is changed into transparent using dimethylbenzene, with canada balsam mountant pair
Section after each histocyte is changed into transparent carries out mounting processing；Bat is observed in section after mounting processing under an optical microscope
According to obtaining and preserve above-mentioned poplar sample cross section micro-image.
3) image pixel is carried out to the poplar micro-image of acquisition and actual size proportionate relationship determines and image procossing, with
Ensure the readability and the accuracy of identification and detection of vessel cell morphological feature in micro-image, specifically include：
3.1) image pixel and actual size dependency relation ratio are determined：The poplar micro-image obtained through step 2) is led to
IMAGE J image analysis programs opening is crossed, the straight line tool at application widget draws engineer's scale length on one and image just
Straight line, the pixel distance value of the straight line can be automatically obtained, then engineer's scale actual (tube) length angle value be inputted, you can determine the micrograph
As image pixel and actual size proportionate relationship, while long measure is set as " μm ", the vessel cell shape obtained during detection
State data will be with " μm " for unit；
3.2) contrast enhancement processing：Adjusted by IMAGE J image analysis programs and determine image pixel through step 3.1)
With the image saturated pixel value of the micro-image of the poplar sample of actual size dependency relation ratio, strengthen and led in the micro-image
The contrast of solencyte and fibrocyte and other cell types, the image saturated pixel pixel value of selection is 0.6, can after processing
Obtain vessel cell and other cell types contrast obvious image；
3.3) image type conversion process：By IMAGE J image analysis programs will be through step 3.2) contrast enhancing at
Image after reason is converted into 8-bit binary system gray level images, i.e. the only image of two kinds of hue distinguishes of black and white, such as Fig. 2 (a) institutes
Show；
3.4) ductal cell marker is handled：The binary system ash for being obtained step 3.3) by IMAGE J image analysis programs
Vessel cell in binary system gray level image is marked by selected threshold for degree image, the optimal threshold of poplar sample micro-image
Value scope is that 100-255, the now chamber of its vessel cell and wall colour all red, completes ductal cell marker processing,
It is converted into (figure Oxford gray represents labeled vessel cell) shown in achromatic image such as Fig. 3 (a)；
4) the micro-image vessel cell morphological feature detection parameters after setting processing, are specifically included：
4.1) vessel cell area parameters are set：Using IMAGE J particle analysis procedure selections after step 3) processing
Larger vessel cell and compared with ductule cell in micro-image, measure respectively after its diameter d according to formula π × (d/2)2Substantially count
Calculate in micro-image larger vessel cell area and compared with ductule cell area, according to poplar sample cross section conduit size,
It is [400,2000] finally to determine its vessel cell areal extent numerical value, and this setting can be by vessel cell area in 400-2000
μm2In the range of vessel cell detect；
4.2) vessel cell circularity parameter setting：It is thin according to conduit to be detected in the micro-image after step 3) processing
The circularity form of born of the same parents sets the vessel cell circularity parameter to be checked measured, because conduit form is more irregular in poplar sample, because
It is 0.3-1 that this, which selects its circularity parameter, wherein, circularity parameter is bigger, conduit form is represented closer to circle, wherein 1 represents rule
Circle then.
5) vessel cell morphological feature detects：Above-mentioned vessel cell morphological feature inspection is completed to poplar sample micro-image
After surveying parameter setting, meet that the conduit of detection parameters requirement is thin by performing IMAGE J graphical analysis particle programs and can obtain
Born of the same parents' image and vessel cell characteristic pattern data, detection select OUTLINES, DISPLAY SESULTS when exporting,
SUMMARIZE and IN SITU SHOW carry out output result form setting, the poplar sample cross section conduit that final detection obtains
Form extracts micro-image as shown in Fig. 4 (a), and vessel cell intermediate record has vessel cell numbering (i.e. in figure in Fig. 4 (a)
Stain among each vessel cell of signal, can see its concrete numerical value clearly after amplification)；The poplar wood vessel cell sum of acquisition
Amount, the gross area, average area, average roundness and all vessel cells detected area shared in whole micro-image
The morphological data overall information such as ratio is as shown in table 1；Single vessel cell numbering and corresponding size and circularity form letter
Breath is as shown in table 2, has been finally completed the detection of poplar vessel cell morphological feature.
Table 1：Vessel cell area and circularity overall numerical value in poplar material
Table 2：Single vessel cell area and circularity numerical value in poplar material
Embodiment 2
1) to obtain Mao bamboo timber micro-image, sample preparation early stage is carried out to Mao bamboo timber：
According to lignocellulosic material Mao bamboo timber shape, mao bamboon stem ring of the cross-sectional area more than 10mm is cut into 10mm (radial direction)
× 10mm (tangential) × 15mm (longitudinal direction) rectangular-shape material.The bulk material obtains as sample for its micro-image.
2) Mao bamboo timber sample can be told by, which obtaining the sample prepared by light microscope, respectively forms the micro- of cell
Image：
The sample that step 1) is prepared, except section statining uses 2% sarranine alcohol dyestuff to dye section, contaminate
The color time is outside 2-4 hours, and other operations are implemented same as Example 1.
3) image pixel is carried out to the Mao bamboo timber micro-image of acquisition and actual size proportionate relationship determines and processing, to protect
The readability and the accuracy of identification and detection of vessel cell morphological feature in micro-image are demonstrate,proved, is specifically included：
3.1) image pixel and actual size dependency relation ratio are determined：It is same as Example 1；
3.2) contrast enhancement processing：It is same as Example 1；
3.3) image type conversion process：It is same as Example 1, obtain the binary system gray-scale map in Mao bamboo timber sample cross section
As shown in Fig. 2 (b)；
3.4) ductal cell marker is handled：The optimal threshold scope of Mao bamboo timber sample micro-image is 170-255, Qi Tacao
Putting into effect, it is same as Example 1 to apply, and achromatic image is converted into after mark, and (figure Oxford gray is represented being labeled as shown in Fig. 3 (b)
Vessel cell).
4) the Mao bamboo timber sample micro-image vessel cell morphological feature detection parameters after setting processing, are specifically included：
4.1) vessel cell area parameters are set：It is final determine Mao bamboo timber vessel cell areal extent numerical value for [8000,
20000]；This setting can be by vessel cell area at 8000-20000 μm2In the range of vessel cell detect, Qi Tacao
Putting into effect, it is same as Example 1 to apply；
4.2) vessel cell circularity parameter setting：Its circularity parameter of the Mao bamboo timber sample of selection is 0.5-1, and other operations are real
Apply same as Example 1；
5) vessel cell morphological feature detects：Detection process is same as Example 1, the Mao bamboo timber sample that final detection obtains
Cross section conduit Morphology observation extracts micro-image as shown in Fig. 4 (b), and vessel cell intermediate record has conduit thin in Fig. 4 (b)
Born of the same parents number (stain among each vessel cell illustrated in figure, can see its concrete numerical value after amplification clearly)；The willow wood of acquisition
Material vessel cell total quantity, the gross area, average area, average roundness and all vessel cells detected are in whole micrograph
The morphological data overall information such as shared area ratio is as shown in table 3 as in；Single vessel cell numbering and corresponding area are big
Small and circularity shape information is as shown in table 4, has been finally completed the detection of Mao bamboo timber vessel cell morphological feature.
Table 3：Conduit area and circularity overall numerical value in mao bamboon material
Table 4：Single conduit area and circularity numerical value in mao bamboon material
Embodiment 3
1) to obtain herba fibraureae recisae material micro-image, sample preparation early stage is carried out to herba fibraureae recisae material：
The herba fibraureae recisae material sample shape of interception is the same as embodiment 2.The bulk material obtains as sample for its micro-image.
2) herba fibraureae recisae material sample can be told by, which obtaining the herba fibraureae recisae material sample prepared by light microscope, respectively forms cell
Micro-image：Herba fibraureae recisae material micro-image obtaining step is the same as embodiment 2.
3) image pixel is carried out to the herba fibraureae recisae material micro-image of acquisition and actual size proportionate relationship determines and processing, to protect
The readability and the accuracy of identification and detection of vessel cell morphological feature in micro-image are demonstrate,proved, is specifically included：
3.1) image pixel and actual size dependency relation ratio are determined：It is same as Example 1；
3.2) contrast enhancement processing：It is same as Example 1；
3.3) image type conversion process：It is same as Example 1, obtain the binary system gray level image in herba fibraureae recisae material cross section such as
Shown in Fig. 2 (c)；
3.4) ductal cell marker is handled：The optimal threshold scope of herba fibraureae recisae material sample micro-image is 153-255, Qi Tacao
Putting into effect, it is same as Example 1 to apply, and achromatic image is converted into after mark, and (figure Oxford gray is represented being labeled as shown in Fig. 3 (c)
Vessel cell).
4) the herba fibraureae recisae material sample micro-image vessel cell morphological feature detection parameters after setting processing, are specifically included：
4.1) vessel cell area parameters are set：It is final determine herba fibraureae recisae material vessel cell areal extent numerical value for [20000,
50000]；This setting can be by vessel cell area at 20000-50000 μm2In the range of vessel cell detect, it is other
Operation is implemented same as Example 1；
4.2) vessel cell circularity parameter setting：Its circularity parameter of the herba fibraureae recisae material sample of selection is 0.8-1, and other operations are real
Apply same as Example 1；
5) vessel cell morphological feature detects：Detection process is same as Example 1, the herba fibraureae recisae material sample that final detection obtains
Cross section conduit Morphology observation extracts micro-image as shown in Fig. 4 (c), and vessel cell intermediate record has conduit thin in Fig. 4 (c)
Born of the same parents number (stain among each vessel cell illustrated in figure, can see its concrete numerical value after amplification clearly)；The herba fibraureae recisae material of acquisition
Vessel cell total quantity, the gross area, average area, average roundness and all vessel cells detected are in whole micro-image
In the morphological data overall information such as shared area ratio it is as shown in table 5；Single vessel cell numbering and corresponding size
It is as shown in table 6 with circularity shape information, it has been finally completed the detection of herba fibraureae recisae material vessel cell morphological feature.
Table 5：Conduit area and circularity overall numerical value in herba fibraureae recisae material
Table 6：Single conduit area and circularity numerical value in herba fibraureae recisae material
Claims (2)
1. a kind of method of vessel cell morphological feature in quick detection lignocellulosic material, it is characterised in that this method is first
It is to obtain micro-image, sample preparation early stage is carried out to lignocellulosic material；The sample prepared is passed through into light microscope
Acquisition can tell the micro-image that each lignocellulosic material respectively forms cell；The micro-image of acquisition is carried out image pixel and
Actual size proportionate relationship determines and processing, to ensure the readability of vessel cell morphological feature and identification and inspection in micro-image
The accuracy of survey；The parameter of micro-image vessel cell morphological feature detection after setting processing；It is special to carry out vessel cell form
Sign detection, it is final to obtain vessel cell image and vessel cell form spy in the lignocellulosic material for meeting setup parameter requirement
Levy data.
2. according to the method for claim 1, it is characterised in that this method specifically includes following steps：
1) to obtain micro-image, sample preparation early stage is carried out to lignocellulosic material：
According to lignocellulosic material species and shape, it is cut into beneficial to the block-shape morphology material for obtaining micro-image, the block material
Material obtains as sample for its micro-image；
2) each lignocellulosic material sample can be told by, which obtaining the sample prepared by light microscope, respectively forms cell
Micro-image：
The sample that step 1) is prepared cuts the section that thickness is 10-30 μm；Then section is contaminated with biological agent dyestuff
Color, to guarantee to distinguish the different types of each histocyte for forming lignocellulosic material, dyeing time foundation shows in optics
Depending on each histiocytic clear-cut required duration observed under micro mirror；Then in being cut into slices by graded ethanol
Each tissue cell dehydration, and each histocyte after dehydration is changed into transparent using dimethylbenzene, with mountant to each histocyte
Section after being changed into transparent carries out mounting processing；Section after mounting processing is observed under an optical microscope takes pictures, and obtains wooden
The micro-image of fibrous material sample, for all types of lignocellulosic material samples, the sample can be distinguished by preserving each sample
The micro-image of middle vessel cell；
3) image pixel is carried out to the micro-image of acquisition and actual size proportionate relationship determines and image procossing, it is micro- to ensure
The readability and the accuracy of identification and detection of vessel cell morphological feature, are specifically included in image：
3.1) image pixel and actual size dependency relation ratio are determined：The engineer's scale on micro-image that will be obtained through step 2)
Physical length it is corresponding with the pixel of the engineer's scale, you can determine the proportionate relationship between image pixel and actual size, be used for
It is the actual size of conduit to ensure what is obtained when detecting vessel cell morphological feature；
3.2) contrast enhancement processing：By adjusting image pixel and actual size dependency relation ratio are determined through step 3.1)
The image saturated pixel value of the micro-image of lignocellulosic material sample, strengthens vessel cell and fibrocyte in the micro-image
And the contrast of other cell types, contribute to the identification of vessel cell morphological feature；
3.3) image type conversion process：Image after step 3.2) contrast enhancement processing is converted into binary system gray-scale map
Picture, the i.e. only image of two kinds of hue distinguishes of black and white；
3.4) ductal cell marker is handled：By the binary system gray level image that step 3.3) obtains by selected threshold to binary system ash
Vessel cell is marked in degree image, is set as most when all designated colors of the only chamber of vessel cell and wall color
Vessel cell in the image has been carried out mark processing by good threshold value, the micro-image obtained by the optimal threshold, this
Mark is easy to the detection subsequent to vessel cell；
4) the micro-image vessel cell morphological feature detection parameters after setting processing, are specifically included：
4.1) vessel cell area parameters are set：Select larger vessel cell and smaller in the micro-image after step 3) processing
Vessel cell, its diameter is measured respectively, and calculate the area S ' of larger vessel cell in fibre imagemaxWith compared with ductule cell
Area S 'min；The vessel cell area lower limit S of settingmin＜ S 'min, the vessel cell area higher limit S of settingmax＞
S’max, and the two round numbers；The vessel cell area S to be detected then set meets：Smin≤S≤Smax；
4.2) vessel cell circularity parameter setting：According to vessel cell to be detected in the micro-image after step 3) processing
The circularity setting vessel cell circularity parameter c to be checked measured, 0 ＜ c≤1, wherein, 1 represents the circle of rule；
5) vessel cell morphological feature detects：After vessel cell morphological feature detection parameters according to step 4) setting, expired
The vessel cell image and vessel cell characteristic pattern data of sufficient detection parameters requirement；The vessel cell characteristic pattern data
Including：Vessel cell morphological data overall information, single vessel cell numbering and corresponding size and circularity shape information,
It is finally completed the detection of lignocellulosic material vessel cell morphological feature.
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