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Detecting method for cell die

A detection method and cell technology, which is applied in the field of judging cell apoptosis, can solve the problems that the detection method cannot be carried out on the same cell sample and affects the accuracy of the detection results, etc.

Inactive Publication Date: 2005-01-05
GUANGXI UNIV
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  • Abstract
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AI Technical Summary

Problems solved by technology

[0006] Although the combination of ordinary light microscopy and TUNEL method is better than single method to detect apoptosis, due to the limitation of various factors, the two detection methods cannot be carried out on the same cell sample, and can only be performed on two adjacent cells. Two methods were used to detect each cell sample, which directly affected the accuracy of the detection results.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] 1. For the buffalo ovary tissue section, according to HE staining requirements, use hematoxylin and eosin for routine staining, observe and take pictures with a light microscope at 400 times. Pink, the nuclei are stained purple-black, consistent with the characteristics of apoptosis.

[0014] 2. Seal the slice with gum. One month later, soak the slice in xylene for 24 hours, then gently remove the cover glass, and then drop xylene to clean the gum on the slice.

[0015] 3. Soak in 100% absolute ethanol three times, 3 minutes each time, to remove xylene.

[0016] 4. Soak eosin in 95%, 90%, 80%, 75%, 70%, and 50% ethanol for 2-3 minutes each time, and finally rinse with double-distilled water for 2-3 minutes. The red color disappears.

[0017] 5. Gently add 75% ethanol solution containing 1% hydrochloric acid (that is, sour alcohol) dropwise to the sliced ​​tissue that is placed horizontally and removes eosin, and covers the tissue for 1-2 minutes, then pour off the sou...

Embodiment 2

[0021] 1. Get the cultured follicular granulosa cells to make a cell smear, and the HE staining method is the same as in Example 1.

[0022] 2. Put the cell smear just sealed on the same day into xylene and soak for 2 hours, remove the gum, and then use the same steps as steps 3, 4, 5, and 6 in Example 1 for detection.

[0023] Apoptosis was confirmed by the results of the two methods.

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Abstract

The invention is a detecting method for cell death; it carries on observation and photo taking to the normal HE dyed cell sample, then uses dimethyl benzene to immerse it to eliminate the tree gum or cedar oil for flaking, then uses waterless alcohol to immerse it to eliminate dimethyl benzene; then uses alcohol with reduced density and the double distilled water to immerse in order to eliminate the eosin in order, then drops in acid alcohol and double distilled water to decolorize the hematine; finally, uses the TUNEL method to detect the cell sample. Then, the method synthesizes the two detecting results, works out the death diagnosis. The invention can realize the detection with two methods on the same cell sample; it can represent the death state of the cell truly and accurately.

Description

technical field [0001] The invention belongs to the technical field of judging cell apoptosis. technical background [0002] There are two ways of cell death, necrosis and apoptosis, which are completely different in morphology and mechanism. Apoptosis is one of the hot topics of research in biology, medicine and other fields in recent years. Since apoptosis is cell death caused by the orderly degradation of cellular DNA under the control of genes, it is also called programmed cell death. Apoptosis is a physiological phenomenon that exists during the development of biological organisms. Once abnormal cell apoptosis occurs, it will lead to diseases in biological organisms, such as tumor formation, immunodeficiency diseases such as AIDS, infectious anemia, etc. Therefore, the detection of apoptosis is of great significance in the occurrence, development and treatment of diseases. There are many methods to study cell apoptosis, such as cell morp...

Claims

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Application Information

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IPC IPC(8): G01N21/25
Inventor 何宝祥杜玉兰
Owner GUANGXI UNIV
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