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Method for observing same cellular morphology by ordinary optical, fluorescence and scanning electron microscopes

A technology of optical microscopy and fluorescence microscopy, which is applied in material analysis, fluorescence/phosphorescence, and scientific instruments through optical means, and can solve problems such as the inability to obtain the three-dimensional outline of cells, the inability to see tissue staining and fluorescence microscopy results in paraffin sections, etc.

Active Publication Date: 2018-11-30
NORTHWEST A & F UNIV
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Problems solved by technology

However, if the wax strip is dewaxed, only the scanned image of the section can be obtained, and the tissue staining and fluorescence microscopy results of the paraffin section cannot be seen; at the same time, the three-dimensional outline of the cells cannot be obtained because the sample has been sectioned

Method used

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  • Method for observing same cellular morphology by ordinary optical, fluorescence and scanning electron microscopes
  • Method for observing same cellular morphology by ordinary optical, fluorescence and scanning electron microscopes
  • Method for observing same cellular morphology by ordinary optical, fluorescence and scanning electron microscopes

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Embodiment 1

[0026] The materials are magnolia ovules and arborvitae ovules

[0027] 1) Fixation: use FAA fixative solution (70% alcohol: formaldehyde: glacial acetic acid = 90:5:5), fix at room temperature for 24 hours.

[0028] 2) Soak in dehydration series solution, soak each reagent for one day, follow the order of Table 1-6.

[0029] Table 1

[0030]

[0031] 3) Dip wax

[0032] Adjust the temperature of the incubator to 60°C, put the sample in a glass bottle, add an appropriate amount of wax shavings, a small amount of tert-butanol, cover it, open the cover two days later, let the tert-butanol volatilize, and soak in the wax for a week.

[0033] 4) Embedding

[0034] Pour the liquid wax into the aluminum box and place it on the spreading machine to prevent solidification. The temperature of the spreading machine is set at 60°C. Put the material soaked in wax for a week into the liquid wax, and use hot tweezers when slowly solidifying at room temperature Stir up and down so tha...

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Abstract

The invention discloses a method for observing same cellular morphology by ordinary optical, fluorescence and scanning electron microscopes. The method is characterized by embedding a sample with paraffin, slicing the tissue of the embedded sample, and determining the slicing progress according to preliminary observation; using the microscopes to observe obtained slices by dyeing of the tissue with a fluorescent composite dye under excitation of white light and different bands of fluorescence to obtain observation results; treating the remaining embedded sample and observing by the scanning electron microscope to obtain a sample section scanning result. The basic morphology, size and basic structure of a cell can be obtained by the ordinary optical microscope, some special structure of thecell can be obtained by the fluorescence microscope, and a three-dimensional image of the interior of the cell can be obtained by the scanning electron microscope. The method is very helpful for thestudy of cell morphology development, as well as the cell internal structure, especially cell nucleus morphology and function.

Description

technical field [0001] The invention belongs to the technical field of biological experiments, and in particular relates to a method for observing three forms of the same cell, namely, an ordinary optical microscope, a fluorescent microscope and a scanning electron microscope. Background technique [0002] Cells are the basic structural and functional units of organisms, and research on cells is a tradition and a hot spot in life sciences. However, for the same cell, the usual method is to observe with an optical microscope; since modern times, transmission electron microscopy and scanning electron microscopy have been used for observation. The transmission electron microscope can observe the internal structure of the cell, and the scanning electron microscope can observe the morphology of the cell surface; The current advanced technology is to use freeze fracture or laser confocal microscopy to observe the internal structure of cells. However, all these microscopes can onl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/84G01N21/64G01N23/2251G01N23/2202
CPCG01N21/6458G01N21/84G01N23/2202G01N23/2251
Inventor 张鑫赵忠李登武裴国亮张国云匡顺贾智硕罗美青
Owner NORTHWEST A & F UNIV
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