CN106525541B - A kind of detection method of fresh cordyceps sinensis cell viability - Google Patents
A kind of detection method of fresh cordyceps sinensis cell viability Download PDFInfo
- Publication number
- CN106525541B CN106525541B CN201610834538.1A CN201610834538A CN106525541B CN 106525541 B CN106525541 B CN 106525541B CN 201610834538 A CN201610834538 A CN 201610834538A CN 106525541 B CN106525541 B CN 106525541B
- Authority
- CN
- China
- Prior art keywords
- cordyceps sinensis
- fresh cordyceps
- dyeing
- cell viability
- slice
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 241001248610 Ophiocordyceps sinensis Species 0.000 title claims abstract description 96
- 230000003833 cell viability Effects 0.000 title claims abstract description 36
- 238000001514 detection method Methods 0.000 title claims abstract description 16
- 238000004043 dyeing Methods 0.000 claims abstract description 57
- 238000000034 method Methods 0.000 claims abstract description 27
- 238000002203 pretreatment Methods 0.000 claims abstract description 19
- 239000000243 solution Substances 0.000 claims description 28
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 7
- 239000008103 glucose Substances 0.000 claims description 7
- 239000008363 phosphate buffer Substances 0.000 claims description 2
- 239000012192 staining solution Substances 0.000 claims description 2
- LCJINPXTMHVZGV-UHFFFAOYSA-N 2,3,5-triphenyl-1h-tetrazol-4-ium;chloride Chemical compound [Cl-].[NH2+]1N(C=2C=CC=CC=2)N(C=2C=CC=CC=2)N=C1C1=CC=CC=C1 LCJINPXTMHVZGV-UHFFFAOYSA-N 0.000 claims 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 3
- 238000010186 staining Methods 0.000 abstract 1
- 241000736199 Paeonia Species 0.000 description 7
- 235000006484 Paeonia officinalis Nutrition 0.000 description 7
- 239000000975 dye Substances 0.000 description 6
- 210000002683 foot Anatomy 0.000 description 6
- 150000003536 tetrazoles Chemical class 0.000 description 5
- 238000003026 viability measurement method Methods 0.000 description 5
- 230000035945 sensitivity Effects 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 3
- 239000001257 hydrogen Substances 0.000 description 3
- 229910052739 hydrogen Inorganic materials 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 244000283482 Alloteropsis cimicina Species 0.000 description 2
- 241000244987 Daiswa polyphylla Species 0.000 description 2
- 101710088194 Dehydrogenase Proteins 0.000 description 2
- 235000017898 Digitaria ciliaris Nutrition 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- VMGAPWLDMVPYIA-HIDZBRGKSA-N n'-amino-n-iminomethanimidamide Chemical compound N\N=C\N=N VMGAPWLDMVPYIA-HIDZBRGKSA-N 0.000 description 2
- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 description 2
- 230000000241 respiratory effect Effects 0.000 description 2
- 230000029058 respiratory gaseous exchange Effects 0.000 description 2
- 101100008048 Caenorhabditis elegans cut-4 gene Proteins 0.000 description 1
- 101100117236 Drosophila melanogaster speck gene Proteins 0.000 description 1
- 244000061458 Solanum melongena Species 0.000 description 1
- 235000002597 Solanum melongena Nutrition 0.000 description 1
- 240000006698 Spigelia anthelmia Species 0.000 description 1
- MOFINMJRLYEONQ-UHFFFAOYSA-N [N].C=1C=CNC=1 Chemical class [N].C=1C=CNC=1 MOFINMJRLYEONQ-UHFFFAOYSA-N 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 238000005138 cryopreservation Methods 0.000 description 1
- 238000005034 decoration Methods 0.000 description 1
- 238000002972 dehydrogenase activity determination Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000007447 staining method Methods 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The present invention relates to detection of agricultural products fields, and in particular to a kind of detection method method of fresh cordyceps sinensis cell viability.Fresh cordyceps sinensis is mainly sliced by this method after pre-treatment, then slice is placed in certain density 2,3, it is impregnated in the small beaker of 5- benzyltriphenylphosphonium chloride tetrazole solution, it is protected from light water bath with thermostatic control dyeing a period of time, the fresh cordyceps sinensis slice after taking out dyeing blots surface moisture, the staining conditions for observing fresh cordyceps sinensis slice, the cell viability of fresh cordyceps sinensis is judged according to the uniformity of the depth of dyeing and dyeing.The present invention is a kind of quick, convenient, method for accurately detecting fresh cordyceps sinensis cell viability.
Description
Technical field
The present invention relates to detection of agricultural products fields, and in particular to a kind of fresh cordyceps sinensis cell of detection quickly, easy is living
The method of power.
Technical background
Cordyceps sinensis is traditional rare traditional Chinese medicine, is existed mostly in the form of its dry product in the market.Due to the fresh winter worm summer
Grass keeps ecosystem, maintains cell viability well, also preferably saves its nutriment and functional component, especially living
The property macromoleculars such as polypeptide and polysaccharide.Therefore, the appearance of instant fresh cordyceps sinensis has been expedited the emergence of in recent years.The freshness of cordyceps sinensis
Very big relationship is maintained with its fresh and crisp mouthfeel and nutrition, functional component.Therefore, how to judge the fresh journey of fresh cordyceps sinensis
Degree also becomes an important project.The research not detected at present to the freshness of fresh cordyceps sinensis still is reported.
The freshness of cordyceps sinensis can be detected by its cell viability to realize.Tetrazole method (TTC method) is widely applied
In the measurement of seed vitality and germination percentage, cardinal principle is that dehydrogenase will exhale when carrying out respiration using living cells in organism
The hydrogen inhaled on chain is sloughed, and tetrazole is a kind of colourless oxidation state species, is transformed into red San Ben Ji formazan after receiving hydrogen
(TTF), active cordyceps sinensis cell will do it respiratory metabolism, and the hydrogen taken off in Respiratory Metabolism Pathway can will be colourless
2,3, 5-Triphenyltertrazoliumchloride (TTC) be reduced into red formazan (TTF) not soluble in water, cell viability is stronger, generation
The activity of thanking also can be more active, and the red degree being dyed to is also deeper, and dead cell is acted on due to breathing no more, thus cannot
It is colored.Therefore fresh cordyceps sinensis may determine that by the dyeing depth and dye uniformity of observing fresh cordyceps sinensis slice
Cell viability, preferably to carry out the quality control of fresh cordyceps sinensis.
It is time-consuming generally all very using tetrazole method measurement seed vitality dyeing course in previously reported technical literature
It is long, generally need 30 hours or more, this is difficult to meet the requirement quickly detected.Such as Chinese patent application (publication number
CN104686011A) in a kind of tetrazole decoration method for measuring eggplant seed viability, seed dyeing time is 15~40 hours.
TTC method has less document report in terms of Chinese medicine vitality test, and there is also the problems that dyeing time is too long.Such as document
" TTC- dehydrogenase reduction method measures the screening of paris polyphylla cell viability optimal conditions " surveys the paris polyphylla TTC method of cryopreservation
The condition for determining cell viability is optimized, and determines that optimum condition is, uses the TTC solution that pH value is 0.8% for 7.0, concentration,
21h is dyed to experimental material under conditions of temperature is 22 DEG C.Document " ginseng-cell TTC- dehydrogenase activity determination condition it is excellent
Change " ginseng-cell vitality test is studied, show that the optimum condition of ginseng-cell vitality test is TTC solution concentration
0.4%, pH 7.5,25 DEG C of reaction temperature, reaction time 18h.The tetrazole method mentioned in the above technical literature in addition to it is time-consuming too
It is long outer, the detection of cordyceps sinensis cell viability is directly applied to there is also repeatability and stability are poor, and susceptibility is low；Again due to time-consuming
Too long, dyeing temperature is higher than fresh cordyceps sinensis storage temperature, and it is living that these methods cannot accurately be truly reflected cordyceps sinensis cell
The problems such as power.
Summary of the invention
The purpose of the present invention is in view of the drawbacks of the prior art and insufficient, the method for taking slice selects suitable slice
Thickness accelerates the detection fresh cordyceps sinensis cell viability time；And pre-treatment is carried out before detection, make the vigor of fresh cordyceps sinensis
It is maintained, so that true, the reliability of testing result；The present invention is also studied on selection slice position, is found
The sensitivity of different parts is different, selects cordyceps sinensis polypide 4-7 that can reflect cell viability faster to sufficient section, shortens inspection
The time is surveyed, sensitivity is increased；The time of similar detection method in the prior art is all very long, and method provided by the invention contaminates
Just can reflect cell viability within color selection of time 1~2 hour, provide it is a kind of it is quick, convenient, accurately detect fresh cordyceps sinensis cell
Vigor reflects the method for cordyceps sinensis freshness.
To achieve the goals above, the technical solution adopted in the present invention the following steps are included:
(1) fresh cordyceps sinensis is taken out from storage condition, carries out pre-treatment；
(2) the fresh cordyceps sinensis Jing Guo pre-treatment is sliced, slice thickness is 1~2mm；
(3) fresh cordyceps sinensis slice is placed in the 2,3,5- benzyltriphenylphosphonium chloride four equipped with mass fraction 0.3%~0.5%
It is impregnated in the small beaker of nitrogen azoles solution, is placed in 37 DEG C and is protected from light dyeing；
(4) height of fresh cordyceps sinensis cell viability is judged according to the uniformity of the depth of dyeing and dyeing, color is got over
Red, dyeing is more uniform, shows that fresh cordyceps sinensis cell viability is stronger.
Preferably, pre-treating method described in step (1) are as follows: 5% glucose that fresh cordyceps sinensis is immersed in 37 DEG C is molten
Liquid 15min.
Preferably, the slice in step (2) is that cordyceps sinensis polypide 4-7 is sliced sufficient section.
Preferably, 2,3, 5-Triphenyltertrazoliumchloride staining solution mass fraction is 0.3% in step (3).
Preferably, dyeing time is 1~2h in step (3).
Preferably, 2,3, 5-Triphenyltertrazoliumchloride solution described in step (3) is matched with the phosphate buffer of pH 7.5
It makes.
It advantage for present invention and has the active effect that
(1) the slice staining method that the present invention takes, has selected suitable slice thickness, has greatly shortened dyeing time,
Improve the efficiency of detection.
(2) present invention has carried out pre-treatment to fresh cordyceps sinensis, preferably saves fresh cordyceps sinensis cell viability, can
The freshness of sample in true reflection storage condition.
(3) present invention is had found during many experiments by analysis, the different parts dyeing of the fresh cordyceps sinensis of same root
There is biggish difference, has chosen cordyceps sinensis polypide, and be sliced to sufficient section for polypide 4-7, dye, greatly improve
The sensitivity of measuring method, accuracy, stability and reproducibility.
(4) the method for the present invention is easy to operate, low in cost, detects without instrument, is capable of the fresh to sample of quicklook
Degree is judged, is suitable for large-scale promotion and is used.
Detailed description of the invention
Fig. 1 is fresh cordyceps sinensis overall diagram, and cordyceps sinensis polypide has 8 pairs of foots, and pereiopoda 3 is right, and close to head, abdominal foot 4 is right,
Uropodium 1 is right, and wherein A is stroma section, and B is polypide head to the 4th pair of sufficient section, and C is polypide 4-7 to sufficient section, and D is polypide
7th pair of foot is to tail portion section.
Fig. 2 is the fresh cordyceps sinensis cell viability measurement result of different dyeing times, and wherein A is dyeing 0.5 hour in figure
Slice afterwards, B are the slices after dyeing 1 hour, and C is the slice after dyeing 2 hours, and D is the slice after dyeing 4 hours, and E is dye
Slice after color 8 hours.
Fig. 3 is different disposal method to fresh cordyceps sinensis cell viability measurement result, wherein figure A is the winter by pre-treatment
Worm summer grass stained slice, B are the cordyceps sinensis stained slices without pre-treatment.
Fig. 4 is the fresh cordyceps sinensis cell viability measurement result of different parts, wherein figure A is stroma stained slice, B is worm
To the 4th pair of sufficient section stained slice, C is polypide 4-7 to sufficient section stained slice on body head, and D is the 7th pair of foot of polypide to tail portion
Section stained slice.
Fig. 5 be various concentration TTC solution dyeing after fresh cordyceps sinensis cell viability measurement result, wherein figure A be through
The slice of 0.1%TTC solution dyeing, B is the slice dyed through 0.2%TTC solution, and C is cut through what 0.3%TTC solution dyed
Piece, D are the slices dyed through 0.4%TTC solution, and E is the slice dyed through 0.5%TTC solution.
Fig. 6 is the fresh cordyceps sinensis cell viability measurement result of different storage times, wherein figure A is after storage 1 month
Stained slice, B are the stained slices after storing 2 months, and C is the stained slice after storing 3 months, and D is the dye after storing 4 months
Color slice, E are the stained slices after storing 5 months, and F is the stained slice after storing 6 months.
Specific embodiment
The embodiment of the present invention is to illustrate technical solution of the present invention rather than its limitations；Although referring to preferred embodiment
Invention is explained in detail, it should be understood by those ordinary skilled in the art that: still can be to tool of the invention
Body embodiment is modified or some technical features can be equivalently replaced；Without departing from the essence of technical solution of the present invention
Mind should all be covered within the scope of the technical scheme claimed by the invention.
Embodiment 1
(1) fresh cordyceps sinensis is taken out from storage condition, carries out pre-treatment, method are as follows: be placed in and be preheating to 37 DEG C
The submergence of 5% glucose solution, 37 DEG C of water-bath 15min；
(2) the fresh cordyceps sinensis polypide part of step (1) processing is sliced, and being sliced position is the 4-7 pairs of polypide
Sufficient section is switched to 1~2mm thin slice, cuts 10；
(3) fresh cordyceps sinensis slice is placed in the small beaker equipped with 0.3% TTC solution and is impregnated with, be placed in 37 DEG C of water
Dyeing is protected from light in bath；
(4) fresh after taking out 2 dyeing respectively when dyeing time is 0.5 hour, 1 hour, 2 hours, 4 hours, 8 hours
Cordyceps sinensis slice, blots surface moisture, observes color, takes pictures.
As a result as shown in Fig. 2, it can be seen from the results that by five different times dyeing, dyeing 0.5 hour after, worm
Grass slice starts to colour, but uneven color, is in pale red；Slice uniform coloring and color after dyeing 1 hour is very deep, is in
Peony；Peony is also presented in color after dyeing 2 hours；The red that dyeing is sliced after 4 hours is thin out, contaminates by 8 hours
After color, the slice color of dyeing is in pale red, or even has white dot.The above result shows that prolonged dyeing can fade, most
Good dyeing time is 1~2 hour.
Embodiment 2
(1) fresh cordyceps sinensis is taken out 2 from storage condition, a progress pre-treatment, method are as follows: be placed in and be preheating to
37 DEG C of 5% glucose solution submergence, 37 DEG C of water-bath 15min；Other one is not done pre-treatment, directly stand-by；
(2) respectively by the fresh cordyceps sinensis of step (1) processing and do not do the cordyceps sinensis polypide same area of pre-treatment into
Row slice, and being sliced position is polypide 4-7 to sufficient section, is switched to 1~2mm thin slice, every is cut 4 respectively；
(3) 2 fresh cordyceps sinensis slices are respectively placed in the small beaker for the TTC solution for being 0.3% equipped with mass fraction
In be impregnated with, be placed in 37 DEG C of water-baths and be protected from light dyeing；
(4) slice for taking out 2 fresh cordyceps sinensis respectively when dyeing time is 2 hours, blots surface moisture, observes face
Color is taken pictures.
As a result as shown in figure 3, it can be seen from the results that by pre-treatment fresh cordyceps sinensis slice color be in peony,
Slice profile all dyes, relatively shallower without the fresh cordyceps sinensis slice profile red of pre-treatment, and section dyeing is not
Completely, there is white dot.The above result shows that can preferably keep the fresh winter worm summer taken out from storage condition by pre-treatment
Careless cell viability, to more really and accurately reflect fresh cordyceps sinensis sample cell viability and freshness.
Embodiment 3
(1) fresh cordyceps sinensis is taken out from storage condition, carries out pre-treatment, method are as follows: be placed in and be preheating to 37 DEG C
The submergence of 5% glucose solution, 37 DEG C of water-bath 15min；
(2) the fresh cordyceps sinensis that step (1) is handled is divided into 4 sections to be sliced, respectively cordyceps sinensis stroma area
Section, polypide head to the 4th pair of sufficient section, 4-7 are to sufficient section and the 7th pair of foot to tail portion, and each section takes 4 in 4 sections,
Every is cut into 1~2mm thin slice；
(3) the fresh cordyceps sinensis slice of 4 different sections is respectively placed in molten for 0.3% TTC equipped with mass fraction
It is impregnated in the small beaker of liquid, is placed in 37 DEG C of water-baths and is protected from light dyeing；
(4) fresh cordyceps sinensis slice is taken out after dyeing 2 hours, is blotted surface moisture, is observed color, take pictures.
As a result as shown in figure 4, centre fails to contaminate it can be seen from the results that stroma section only has edge in light red
Color, polypide head is very shallow to the 4th pair of sufficient section dyeing, and center has hickie to be unstained, and polypide 4-7 is most deep to the dyeing of sufficient section,
All dyeing is in peony to section, and the 7th pair of foot is shallower to the dyeing of tail portion section, also has a small amount of hickie to be unstained.Result above table
The fresh cordyceps sinensis cell viability of embodiment bright, that polypide 4-7 can be best to sufficient section, improves the sensitivity of detection, thus more
Add and really and accurately reflects fresh cordyceps sinensis sample cell viability and freshness.
Embodiment 4
(1) fresh cordyceps sinensis is taken out to a progress pre-treatment, method are as follows: be placed in and be preheating to 37 DEG C from storage condition
5% glucose solution submergence, 37 DEG C of water-bath 15min；
(2) the fresh cordyceps sinensis that step (1) is handled is sliced, and being sliced position is polypide 4-7 to sufficient section, is cut
To 1~2mm thin slice, 10 are cut；
(3) 10 fresh cordyceps sinensis slices are divided into 5 groups, being respectively put into five equipped with appropriate mass fraction is 0.1%,
It is impregnated in the small beaker of 0.2%, 0.3%, 0.4%, 0.5%TTC solution, is placed in 37 DEG C of water-baths and is protected from light dyeing 2 hours；
(4) slice that fresh cordyceps sinensis is taken out after dyeing time is 2 hours, blots surface moisture, observes color, clap
According to.
As a result as shown in figure 5, it can be seen from the results that through mass fraction be 0.1% and 0.2% TTC solution dye
Slice profile red is relatively shallower, and section dyeing is not exclusively, there is a small amount of white dot, and be 0.3% through mass fraction,
The slice color of 0.4% and 0.5% TTC solution dyeing is uniform, and is in peony, the TTC concentration for being 0.3% with mass fraction
More preferably.The above result shows that can preferably be detected using the TTC solution that mass fraction is 0.3%-0.5% from storage condition
The fresh cordyceps sinensis cell viability taken out, to more really and accurately reflect fresh cordyceps sinensis sample cell viability and fresh
Degree.
Embodiment 5
(1) take by different storage times fresh cordyceps sinensis 6 (storage time is respectively 1 month, 2 months, 3 months,
4 months, 5 months, 6 months), carry out pre-treatment, method are as follows: be placed in be preheating to 37 DEG C 5% glucose solution submergence, 37 DEG C
Water-bath 15min；
(2) the fresh cordyceps sinensis of 6 different storage times of step (1) processing is sliced, and be sliced position be away from
From polypide 4-7 to sufficient section, it is switched to 1~2mm thin slice, every fresh cordyceps sinensis cuts 4 respectively；
(3) 6 fresh cordyceps sinensis slices are respectively placed in the small beaker for the TTC solution for being 0.3% equipped with mass fraction
In be impregnated with, be placed in 37 DEG C of water-baths and be protected from light dyeing；
(4) slice that fresh cordyceps sinensis is taken out after dyeing time is 2 hours, blots surface moisture, observes color, clap
According to.
As a result as shown in fig. 6, it can be seen from the results that the fresh cordyceps sinensis dyeing of 6 different storage times has significantly
Difference, wherein the fresh cordyceps sinensis of storage 1 month is dyed in peony, and section even dyeing, without white dot；Storage 2
Fresh cordyceps sinensis dyeing in a month is in peony, but section has a small amount of white dot, speck area very little；3 months fresh of storage
Cordyceps sinensis dyeing is in light red, and section has a small amount of white dot slightly more；The fresh cordyceps sinensis dyeing of storage 4 months is in pale red
Color, section white patch, area are larger；Only outer rim small area is in light red, centre base to the fresh cordyceps sinensis of storage 5 months
Originally it is unstained；The fresh cordyceps sinensis of storage 6 months is all unstained, and section is white.The above result shows that the method for the present invention energy
Enough cell viabilities for detecting fresh cordyceps sinensis quickly, easy, judge the substantially storage time of fresh cordyceps sinensis.
Claims (4)
1. a kind of detection method of fresh cordyceps sinensis cell viability, which comprises the following steps:
(1) fresh cordyceps sinensis is taken out from storage condition, carries out pre-treatment；Pre-treating method are as follows: submerge fresh cordyceps sinensis
In 37 DEG C of 5% glucose solution 15min；
(2) the fresh cordyceps sinensis Jing Guo pre-treatment is sliced, slice thickness is 1~2mm；Slice is cordyceps sinensis polypide the
4-7 is sliced sufficient section；
(3) fresh cordyceps sinensis slice is placed in the 2,3,5 triphenyltetrazolium chlorid equipped with mass fraction 0.3%~0.5%
It is impregnated in the small beaker of solution, is placed in 37 DEG C and is protected from light dyeing；
(4) height of fresh cordyceps sinensis cell viability is judged according to the uniformity of the depth of dyeing and dyeing, color is redder, dye
Color is more uniform, shows that fresh cordyceps sinensis cell viability is stronger.
2. the detection method of fresh cordyceps sinensis cell viability according to claim 1, which is characterized in that 2 in step (3),
3,5- benzyltriphenylphosphonium chloride tetrazole staining solution mass fraction is 0.3%.
3. the detection method of fresh cordyceps sinensis cell viability according to claim 1, which is characterized in that dye in step (3)
The color time is 1~2h.
4. the detection method of fresh cordyceps sinensis cell viability according to claim 1, which is characterized in that institute in step (3)
2,3,5 triphenyltetrazolium chlorid solution is stated to be formulated with the phosphate buffer of pH 7.5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610834538.1A CN106525541B (en) | 2016-09-20 | 2016-09-20 | A kind of detection method of fresh cordyceps sinensis cell viability |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610834538.1A CN106525541B (en) | 2016-09-20 | 2016-09-20 | A kind of detection method of fresh cordyceps sinensis cell viability |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106525541A CN106525541A (en) | 2017-03-22 |
| CN106525541B true CN106525541B (en) | 2019-01-18 |
Family
ID=58343932
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610834538.1A Active CN106525541B (en) | 2016-09-20 | 2016-09-20 | A kind of detection method of fresh cordyceps sinensis cell viability |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106525541B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108693180B (en) * | 2018-06-29 | 2023-12-08 | 湖南洪江嵩云禽业有限公司 | Safety warning card for storage and transportation freshness of chilled poultry meat and use method thereof |
| CN110117791B (en) * | 2019-05-23 | 2023-11-10 | 电子科技大学 | Binding force promoter for brown oxide liquid of high-density interconnection printed circuit board |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5420017A (en) * | 1991-01-24 | 1995-05-30 | Orion Corporation Ltd. | Method and kit for the detection of microorganisms |
| CN1598578A (en) * | 2004-07-22 | 2005-03-23 | 哈尔滨工业大学 | Method for investigating TTC-dehydrogenase activity for drinking water treated biologic |
| CN101423802A (en) * | 2008-12-19 | 2009-05-06 | 重庆市中药研究院 | High-activity Chinese Caterpillar Fungus bacteria and oil suspension |
| CN101569255B (en) * | 2009-06-05 | 2011-11-16 | 中国科学院植物研究所 | Method for detecting seed viability of orchid plants |
| JP6184865B2 (en) * | 2010-07-23 | 2017-08-23 | アステラス インスティテュート フォー リジェネレイティブ メディシン | Methods and highly purified compositions for detecting rare subpopulations of cells |
-
2016
- 2016-09-20 CN CN201610834538.1A patent/CN106525541B/en active Active
Non-Patent Citations (2)
| Title |
|---|
| 一种丝状真菌活细胞生物量测定方法的研究;刘伯宁 等;《药物生物技术》;20061231;第13卷(第1期);第40-44页 |
| 蛹虫草TTC-脱氢酶测定方法的优化;黄春花 等;《环境昆虫学报》;20111231;第33卷(第3期);第321-328页 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106525541A (en) | 2017-03-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Lu et al. | New ratiometric optical oxygen and pH dual sensors with three emission colors for measuring photosynthetic activity in cyanobacteria | |
| Eppley | AN INCUBATION METHOD FOR ESTIMATING THE CARBON CONTENT OF PHYTOPLANKTON IN NATURAL SAMPLES 1 | |
| de Beer et al. | A fast‐responding CO2 microelectrode for profiling sediments, microbial mats, and biofilms | |
| CN106525541B (en) | A kind of detection method of fresh cordyceps sinensis cell viability | |
| NO300853B1 (en) | Assay methods, reagent composition and use thereof in glucose determination | |
| CN105671127B (en) | A kind of stable enzyme process serum Mg-ion detection kit | |
| Turkia et al. | Capillary electrophoresis with laser-induced fluorescence detection for studying amino acid uptake by yeast during beer fermentation | |
| CN102043006B (en) | Method for preparing water-soluble quantum dot carbon paste electrode for detecting trace amino acid in food | |
| Azab et al. | High-throughput sensing microtiter plate for determination of biogenic amines in seafood using fluorescence or eye-vision | |
| CN105238848B (en) | A kind of alpha-L-fucosidase detection kit and preparation method | |
| Rivkin | Photoadaptation in marine phytoplankton: Variations in ribulose 1, 5-bisphosphate activity. | |
| Wang et al. | Silk fibroin-based colorimetric microneedle patch for rapid detection of spoilage in packaged salmon samples | |
| Chen et al. | Fast and Effective Turn‐on Paper‐based Phosphorescence Biosensor for Detection of Glucose in Serum | |
| CN102323258B (en) | Detection test paper for detecting benzoyl peroxide in flour | |
| Sochorova et al. | Electrochemical and others techniques for the determination of malic acid and tartaric acid in must and wine | |
| CN103528977A (en) | Serum beta-hydroxybutyric acid reagent kit and assay method thereof | |
| CN105806839A (en) | Embryo-free rice freshness detecting reagent and method | |
| JPS6047695A (en) | Simple method for determination of freshness-judgement constant k of fish and shellfish | |
| CN105572349B (en) | The preparation and its application of the bionical Rapid detection test strip of agricultural chemicals sevin molecular engram | |
| Paranjpe et al. | A disposable optrode using immobilized tyrosinase films | |
| CN108329316B (en) | A kind of biological fluorescent labeling and preparation method thereof detecting nickel ion | |
| Keay et al. | Applications of flow injection analysis to analytical biotechnology | |
| CN106198406A (en) | A kind of method of quick detection high whiteness flour wheat seed | |
| Podrazký et al. | Monitoring the growth and stress responses of yeast cells by two-dimensional fluorescence spectroscopy: first results | |
| CN108693136A (en) | A kind of method of quick detection pollen activity |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| REG | Reference to a national code |
Ref country code: HK Ref legal event code: DE Ref document number: 1230716 Country of ref document: HK |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Method for detecting cell viability of fresh Cordyceps sinensis Effective date of registration: 20190903 Granted publication date: 20190118 Pledgee: Yichang Branch of Bohai Bank Co., Ltd.|Three Gorges Branch of Bank of China Co., Ltd.|Nanhu Branch of Hubei Bank Co., Ltd.|Yichang Branch of Hankou Bank Co., Ltd. Pledgor: Guangdong Dongguang Sunshine Pharmaceutical Co., Ltd.|Yichang Shancheng Shuidu Cordyceps sinensis Co., Ltd. Registration number: Y2019420000010 |
|
| REG | Reference to a national code |
Ref country code: HK Ref legal event code: WD Ref document number: 1230716 Country of ref document: HK |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20210603 Granted publication date: 20190118 Pledgee: Bank of Bohai Limited by Share Ltd. Yichang branch|Bank of China Limited by Share Ltd. Three Gorges Branch|Bank of Hubei Limited by Share Ltd. Yichang Nanhu sub branch|Bank of Hankou Limited by Share Ltd. Yichang branch Pledgor: SUNSHINE LAKE PHARMA Co.,Ltd.|YICHANG SHANCHENGSHUIDU CORDYCEPS Co.,Ltd. Registration number: Y2019420000010 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20210623 Address after: 443300 baotawan village, Lucheng, Yidu City, Yichang City, Hubei Province Patentee after: YICHANG SHANCHENGSHUIDU CORDYCEPS Co.,Ltd. Address before: 523808 No. 1 Industrial North Road, Songshan Industrial Park, Songshan, Guangdong, Dongguan, Hubei Patentee before: SUNSHINE LAKE PHARMA Co.,Ltd. Patentee before: YICHANG SHANCHENGSHUIDU CORDYCEPS Co.,Ltd. |
|
| TR01 | Transfer of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: A method for detecting cell viability of fresh Cordyceps sinensis Effective date of registration: 20210630 Granted publication date: 20190118 Pledgee: Bank of Bohai Limited by Share Ltd. Yichang branch|Bank of China Limited by Share Ltd. Three Gorges Branch|Bank of Hubei Limited by Share Ltd. Yichang Nanhu sub branch|Bank of Hankou Limited by Share Ltd. Yichang branch Pledgor: YICHANG SHANCHENGSHUIDU CORDYCEPS Co.,Ltd. Registration number: Y2021420000062 |