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Preparation method of cell slide

A cell and slide technology, applied in the field of cell slide preparation, can solve problems such as difficult quality control, inability to stack and overlap cells, growth density of slides, etc., and achieve the effect of reliable operation process

Inactive Publication Date: 2014-07-16
CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no reports on the specific and good operating procedures and key technologies for the preparation of cell slides. As a result, researchers need to repeatedly explore this technology in actual operation, and the quality is difficult to control.
Due to the lack of a standardized process reference, when preparing cell slides, the problem of inoculated cell density is often too large or too small, resulting in overlapping cell accumulation and unable to grow normally on the slide, or the density is too low for subsequent detection, and even crawling occurs. The cells on the sheet are easy to come off the sheet during subsequent processing
This undoubtedly greatly wastes the energy and time of scientific researchers, and also wastes scientific research resources, which greatly affects the application of cell slides in cell immunochemical experiments.

Method used

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  • Preparation method of cell slide

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Embodiment 1

[0057] First, gently place 12 wet sterile slides into the bottom of a 12-well culture plate with tweezers. Then the cells in the logarithmic growth phase were taken, digested with preheated 0.25% trypsin, the cells were blown into a single cell suspension, and the cell density was adjusted to 1.25×10 5 About / ml, use a 1ml pipette gun to draw 1ml of the cell suspension, and vertically and quickly dispense the cell suspension at a distance of about 3cm from the bottom of the 12-well plate. 2 Cultivate in the incubator for 24h. After the cells adhered to the wall the next day, discard the culture medium, wash the cells once with PBS, and add serum-free DMEM culture medium, 1ml per well. Place the 12-well plate at 37°C, 5% CO 2 After continuing to culture in the incubator for 24 hours, take it out of the incubator, discard the supernatant, wash the cells with pre-cooled PBS for 3 times, add 1ml of 4% paraformaldehyde to fix for 30min, discard the fixative and then use the pre-c...

Embodiment 2

[0061] Use tweezers to lightly put the wet sterile slide into the bottom of the 12-well plate, then take the cells in the logarithmic growth phase, digest them with preheated 0.25% trypsin, blow the cells into a single-cell suspension, and separate the cells Density adjusted to 1.5×10 5 About / ml, use a 1ml pipette gun to draw 1ml of cell suspension, and vertically and quickly pipette the cell suspension at a distance of about 4cm from the bottom of the 12-well plate. 2 Cultivate in the incubator for 24h. After the cells adhere to the wall the next day, discard the culture medium, wash the cells once with PBS buffer, and add serum-free DMEM culture medium, 1ml per well. Place the 12-well plate at 37°C, 5% CO 2 After continuing to culture in the incubator for 24 hours, take it out of the incubator, discard the supernatant, wash the cells with pre-cooled PBS for 3 times, add 1ml of 4% paraformaldehyde for fixation for 40min, discard the fixative and then use the pre-cooled Wa...

Embodiment 3

[0073] Use tweezers to lightly put the wet sterile slide into the bottom of the 12-well plate, then take the cells in the logarithmic growth phase, digest them with preheated 0.25% trypsin, blow the cells into a single-cell suspension, and separate the cells Density adjusted to 2.0×10 5 About / ml, use a 1ml pipette gun to draw 1ml of cell suspension, and vertically and quickly pipette the cell suspension at a distance of about 5cm from the bottom of the 12-well plate. 2 Cultivate in the incubator for 24h. After the cells adhered to the wall the next day, discard the culture medium, wash the cells once with PBS, and add serum-free DMEM culture medium, 1ml per well. Place the 12-well plate at 37°C, 5% CO 2 After continuing to culture in the incubator for 24 hours, take it out of the incubator, discard the supernatant, wash the cells with pre-cooled PBS for 3 times, add 1ml of 4% paraformaldehyde to fix for 50min, discard the fixative and then use the pre-cooled Wash with cold...

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Abstract

The invention relates to a preparation method of a cell slide. The method comprises the following steps of cutting the slide according to the need, and sterilizing; completely soaking the sterilized slide into a culture solution, and then putting onto a culture plate; sucking cell suspension by a pipette, and vertically spraying out the cell suspension to the part just above the culture plate for once to enable cells to be evenly distributed on the slide under the action of the impact force; adding stationary liquid into the culture plate for fixing; and carrying out chemical staining on immune cells. A good and reliable concrete operation process and key technology improvement scheme is provided for the preparation of the cell slide and the subsequent cellular immunochemistry.

Description

technical field [0001] The invention relates to a method for preparing cell slides. Background technique [0002] Cell climbing technology is a technique that uses the habit of adherent cells to grow on a certain solid surface (such as a cover glass) to perform in vitro experimental operations. When using cell slides for in vitro experiments, the influence of many interfering factors in the body can be eliminated, and various interventions can be conveniently performed on cells. Therefore, the preparation of cell slides is an important technique for in vitro cell culture. At the same time, the preparation of cell slides is the first step in immunocytochemistry, immunofluorescence, TUNEL apoptosis detection, etc., and it is also a key step to successfully obtain experimental results. At present, with the advocacy and implementation of the principle of "3Rs" (replacement, reduction and refinement) and the deepening of research work, in vitro cell research and detection indic...

Claims

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Application Information

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IPC IPC(8): G01N1/28
Inventor 周显青徐英郭芳子李艳博孙志伟
Owner CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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