CN103674980A - Microscopic observation method of diatom - Google Patents
Microscopic observation method of diatom Download PDFInfo
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- CN103674980A CN103674980A CN201310643497.4A CN201310643497A CN103674980A CN 103674980 A CN103674980 A CN 103674980A CN 201310643497 A CN201310643497 A CN 201310643497A CN 103674980 A CN103674980 A CN 103674980A
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Abstract
A microscopic observation method of diatom is characterized in that the method comprises the following steps: putting a diatom specimen under a low-magnification light microscope, carrying out microscopic examination, observing the diatom specimen under a scanning electron microscope, and observing the diatom specimen under a high-magnification light microscope. The microscopic observation method of diatom allows the diatom specimen to be still observed under the light microscope and a clear shell surface pattern to be obtained after the diatom specimen is observed under the scanning electron microscope. The method is especially suitable for rare diatom varieties, improves the specimen utilization rate, shortens the test period, and saves the high use cost of the scanning electron microscope.
Description
Technical field
The present invention relates to biological scientific experiment technical field, specifically just relate to the microexamination of plant specimen, especially relate to a kind of method of diatom microexamination.
Background technology
Diatom is a class Eukaryotic Algae, and kind is many, and quantity is large, and widely distributed.Diatom is primary productivity main in ocean and fresh water, and tool vital role in global carbon and silicon circulation, is more and more applied on monitoring water environment, ancient ecological reconstruction and Relation To Strata simultaneously.Frustule has the cell membrane of height silicidize.By upper and lower two valve jackets, closed and formed, shell mask has symmetria bilateralis or radiosymmetric decorative pattern.Identifying of diatom kind is mainly that to take form and the lip-deep decorative pattern of shell of diatom test be foundation.In order to see the decorative pattern on diatom test clearly, before microexamination, sample must, first through clearing up processing, be removed its inclusions (being mainly organic).
Traditional diatom microscopy observation method comprises that the sample after light Microscopic observation and cancellation solution is observed two steps under scanning electron microscope with permanent mounting at present.Diatom samples, after clearing up, is kept in anhydrous alcohol, shakes up, and draws 20ml sample and is placed on cover glass, after drying, by mounting rubber seal, enters in microslide, after fully air-dry, becomes permanent envelope sheet, can be used for om observation.Once but Diatom samples make after permanent mounting, the Diatom samples in mounting just can not be for scanning electron microscopic observation.When taking scanning electron microscopic observation, one of first clip is bonded on copper platform with copper platform conducting resinl of the same size, the sample 10-15ml that absorption is stored in anhydrous alcohol drops on the copper platform that is stained with conducting resinl, in this step, sample does not pass through light microscopy checking, therefore can not confirm whether to have in sample the kind of needs, rare diatom particularly, the quantity in sample seldom, is difficult to guarantee that each drips in sample and can have.Treat that the sample on copper platform is thoroughly air-dry, general about 1 day, or be positioned in drying baker 20 minutes.Metal spraying; putting into scanning electron microscope observes; the volume of diatom kind is smaller; general length 10-100 μ m; copper platform of electricity Microscopic observation needs about 2 hours, if sample without light microscopy checking, often can not find the kind needing during scanning electron microscopic observation; the energy of so both having lost time, wastes energy again.Current Diatom samples will wash after scanning electron microscopic observation, if there is rare diatom in sample, will be significant wastage to sample.
Summary of the invention
Technical matters object of the present invention is cannot be for the technological deficiency of om observation after scanning electron microscopic observation for existing Diatom samples, a kind of method of diatom microexamination is provided, Diatom samples still can carry out om observation after scanning electron microscopic observation, and can obtain the clear decorative pattern of shell face, the method is particularly useful for the few and rare diatom kind of quantity, can improve the utilization factor of sample, shorten the test period, save the great number cost that uses scanning electron microscope.
Technical scheme
In order to realize above-mentioned technical purpose, the present invention designs a kind of method of diatom microexamination, it is characterized in that, it comprises following step:
(1) Diatom samples is placed on microscopy under low power light microscopic, is about to be placed on slide through clearing up the sample taking-up of processing, at low power Microscopic observation;
(2) Diatom samples is observed under scanning electron microscope, and the sample that is about to cross through light microscopy checking is bonded on copper platform, through metal spraying, puts into scanning electron microscope observation and takes pictures;
(3) Diatom samples is at high power light Microscopic observation, and the sample that soon scanning electron microscopic observation will be crossed takes off from copper platform, makes after permanent mounting, at high power Microscopic observation, takes pictures.
Further, when described step (1) Diatom samples is placed under low power light microscopic microscopy, the described sample of clearing up after processing is taken out, after sample fully need to being vibrated on oscillator during taking-up, with liquid-transfering gun, draw 20～30ml, be ready to clean microslide simultaneously, on microslide, place with the circular lid slide number of copper platform consistent size and open, general 6～8, be evenly arranged on microslide, and aside with marking pen, number.The sample of taking-up is splashed on circular cover plate successively, standing 2 minutes, be placed under 20 times or 40 times of light microscopics and check, by having in sample, need the cover plate of kind number record to get off.
Further, while observing under described step (2) Diatom samples scanning electron microscope, on copper platform, paste 1～2 fine strip shape conducting resinl, the circular cover plate of the good numbering of record is carefully placed on clean copper platform, treats fully air-dryly, or be positioned in drying baker 20 minutes, metal spraying, put into scanning electron microscope and observe, search needed kind and take pictures, record kind quantity and position on each copper platform.
Further, when described step (3) Diatom samples high power light Microscopic observation, first the copper platform after electron microscopic observation is taken out, with tweezers, circular cover plate is taken off from copper platform, with mounting glue, there is the one side of sample to seal on microslide cover plate, treat that mounting glue is completely air-dry, be placed under 100 times of light microscopics and observe with oil dripping, needed kind is carried out to om observation and take pictures, kind quantity and the position of in the time of now can be with reference to electron microscopic observation, recording.
Beneficial effect
The method of a kind of diatom microexamination provided by the invention, make Diatom samples after scanning electron microscopic observation, still can carry out om observation, and can obtain the method for the clear decorative pattern of shell face, the method is specially adapted to the few and rare diatom kind of quantity, improve the utilization factor of sample, can also shorten the test period, save the great number cost that uses scanning electron microscope simultaneously.
Accompanying drawing explanation
Accompanying drawing 1 is process step schematic diagram of the present invention.
Embodiment
Below in conjunction with drawings and Examples, the present invention will be further described,
Embodiment
As shown in Figure 1, the microexamination of Neidium iridis sample of take is example, and it comprises following step:
First Diatom samples is placed on to microscopy under low power light microscopic, be about to be placed on slide through clearing up the sample taking-up of processing, at low power Microscopic observation, be exactly specifically: the described sample of clearing up after processing is taken out, after sample fully need to being vibrated on oscillator during taking-up, with liquid-transfering gun, draw 20～30ml, be ready to clean microslide simultaneously, on microslide, place with the circular lid slide number of copper platform consistent size and open, general 6～8, be evenly arranged on microslide, and aside with marking pen, number.The sample of taking-up is splashed on circular cover plate successively, standing 2 minutes, be placed under 20 times of 40 times of light microscopics and check, by having in sample, need the cover plate of kind number record to get off.
Then under Diatom samples scanning electron microscope, observe, the sample that is about to cross through light microscopy checking is bonded on copper platform, through metal spraying, puts into scanning electron microscope and observes, specifically on copper platform, paste exactly 1～2 fine strip shape conducting resinl, the circular cover plate of the good numbering of record is carefully placed on clean copper platform, treats fully air-dryly, or be positioned in drying baker 20 minutes, metal spraying, put into scanning electron microscope and observe, search needed kind and take pictures, record kind quantity and position on each copper platform.
Last Diatom samples high power light Microscopic observation, the sample that soon scanning electron microscopic observation will be crossed takes off from copper platform, make after permanent mounting, at 100 times of light Microscopic observations, take pictures, be exactly specifically that the copper platform that takes Electronic Speculum is taken out, with tweezers, circular cover plate is taken off from copper platform, with mounting glue, there is the one side of sample to seal on microslide cover plate, treat that mounting glue is completely air-dry, be placed under 100 times of light microscopics and observe with oil dripping, needed kind is carried out to om observation and take pictures, kind quantity and the position of in the time of now can be with reference to electron microscopic observation, recording.
Compared with prior art, advantage of the present invention comprises that under sample low power light microscopic, microscopy, sample are observed and sample high power light Microscopic observation step under scanning electron microscope successively.Under sample low power light microscopic, in microscopy step, the described sample of clearing up after processing is first shaken up with oscillator, can reduce the impurity being attached on cell like this, the more situation of impurity in particularly suitable sample.Sample splashes on ready circular cover plate, is placed under 20 times or 40 times of light microscopics and checks, the cover plate number record of effective sample (being to have needed kind in sample) is got off, and can know like this kind situation on each cover plate.A general cover plate has the diatom of many types and quantity, and some kind is that we are needed, at this moment just the cover plate at kind place is recorded and is kept, and has so just avoided blindness, shortens the test period.Mostly technology is in the past sample directly to drip on copper platform and carry out electron microscopic observation, all there is no the kind of needs in a lot of samples, has so both wasted time and labour, has also wasted the energy.Under sample scanning electron microscope, observe in step, paste～2 fine strip shape conducting resinls on copper platform, scanning electron microscopic observation is just easy to cover plate to take off after complete like this, and through evidence, the fastness of fine strip shape conducting resinl is also fine.Technology is in the past on copper platform, to be all covered with conducting resinl, so both to have wasted conducting resinl, and cover plate is also difficult to take off from Tong Tai.In sample high power light Microscopic observation step, with tweezers, circular cover plate is taken off from copper platform, with mounting glue, sample is sealed on microslide, be placed on 100 times of light Microscopic observations after air-dry.This scanning electron microscope sample is carried out to the method for om observation again, effectively improved the utilization factor of sample, reduced the waste to sample, be specially adapted to the few and rare diatom kind of quantity, in om observation, the record of kind in the time of can be with reference to electron microscopic observation, in case miss some important kind samples.Therefore, the method of a kind of diatom microexamination provided by the invention, make Diatom samples after scanning electron microscopic observation, still can carry out om observation, and can obtain the method for the clear decorative pattern of shell face, the method is specially adapted to the few and rare diatom kind of quantity, improves the utilization factor of sample, can also shorten the test period, save the great number cost that uses scanning electron microscope simultaneously.
Claims (4)
1. a method for diatom microexamination, is characterized in that, it comprises following step:
(1) Diatom samples microscopy under low power light microscopic, is about to be placed on slide through clearing up the sample taking-up of processing, at low power Microscopic observation;
(2) Diatom samples is observed under scanning electron microscope, and the sample that is about to cross through light microscopy checking is bonded on copper platform, through metal spraying, puts into scanning electron microscope observation and takes pictures;
(3) Diatom samples high power light Microscopic observation, the sample that soon scanning electron microscopic observation will be crossed takes off from copper platform, makes after permanent mounting, at high power light Microscopic observation, takes pictures.
2. the method for a kind of diatom microexamination as claimed in claim 1, it is characterized in that: when described step (1) Diatom samples is placed under low power light microscopic microscopy, the described sample of clearing up after processing is taken out, after sample fully need to being vibrated on oscillator during taking-up, with liquid-transfering gun, draw 20～30ml, be ready to clean microslide simultaneously, on microslide, place with the circular lid slide number of copper platform consistent size and open, general 6～8, be evenly arranged on microslide, and aside with marking pen, number.The sample of taking-up is splashed on circular cover plate successively, standing 2 minutes, be placed under 20 times or 40 times of light microscopics and check, by having in sample, need the cover plate of kind number record to get off.
3. the method for a kind of diatom microexamination as claimed in claim 1, it is characterized in that: while observing under described step (2) Diatom samples scanning electron microscope, on copper platform, paste 1～2 fine strip shape conducting resinl, the circular cover plate of the good numbering of record is carefully placed on clean copper platform, treats fully air-dryly, or be positioned in drying baker 20 minutes, metal spraying, put into scanning electron microscope and observe, search needed kind and take pictures, record kind quantity and position on each copper platform.
4. the method for a kind of diatom microexamination as claimed in claim 1, it is characterized in that: when described step (3) Diatom samples high power light Microscopic observation, first the copper platform after electron microscopic observation is taken out, with tweezers, circular cover plate is taken off from copper platform, with mounting glue, there is the one side of sample to seal on microslide cover plate, treat that mounting glue is completely air-dry, being placed on oil dripping under 100 times of light microscopics observes, needed kind is carried out to om observation and take pictures, kind quantity and the position of in the time of now can be with reference to electron microscopic observation, recording.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105784705A (en) * | 2016-02-21 | 2016-07-20 | 山东政法学院 | Forensic medicine diatom test method |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105784705A (en) * | 2016-02-21 | 2016-07-20 | 山东政法学院 | Forensic medicine diatom test method |
| CN105784705B (en) * | 2016-02-21 | 2018-05-18 | 山东政法学院 | A kind of diatom inspection method in medical jurisprudence |
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