A kind of preparation method of biological single cell transmission electron microscope sample
A transmission electron microscope sample and single-cell technology, which is applied in the preparation of test samples, sampling, and material analysis using radiation, etc., can solve the problems of difficult to clearly visualize the ultrastructure of cells and high failure rate, and improve the success rate. Strong experimental compatibility and cost-saving effect
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Embodiment 1
[0027] The specific steps of the method for preparing ultrathin section samples of Chlamydomonas reinhardtii by using lead solution as vital staining agent:
[0028] (1) In vivo staining: Take a sterilized 250ml Erlenmeyer flask, add 100ml TAP liquid medium, transfer 1mL of purified Chlamydomonas reinhardtii culture medium into the medium with a pipette gun, and then transfer it with another pipette gun 50ul of sterile Pb(NO) with a concentration of 1000ug / L 3 ) 2 Solution enters culture medium (medium lead concentration 500ug / L), seals with aseptic parafilm, shakes up;
[0029] (2) Cultivation: the Erlenmeyer flask after step (1) was inoculated was carried out at 25° C. for 12 hours of light and 12 hours of darkness for 4 days;
[0030] (3) Pre-fixation: Take an appropriate amount of Chlamydomonas reinhardtii sample solution, add glutaraldehyde solution with a concentration of 2.5% by volume, and fix for 2 hours at 4°C without light;
[0031] (4) Rinse: wash the sample 4 t...
Embodiment 2
[0038] The specific steps of the method for preparing ultrathin section samples of Chlamydomonas reinhardtii by using arsenic and lead mixed solution as vital staining agent:
[0039] (1) In vivo staining: Take a sterilized 250ml Erlenmeyer flask, add 100ml TAP liquid medium, transfer 1ml of purified Chlamydomonas reinhardtii culture solution into the medium with a pipette gun, and then transfer 50ul with another pipette gun Concentration of 1000mg / L sterile Pb(NO 3 ) 2 Solution and 50ul concentration of 100mg / L arsenic solution standard substance mixture into the culture medium, sealed with sterile parafilm, shake well;
[0040] (2) Cultivation: the Erlenmeyer flask after step (1) was inoculated was carried out at 25° C. for 12 hours of light and 12 hours of darkness for 4 days;
[0041](3) Pre-fixation: Take an appropriate amount of Chlamydomonas reinhardtii sample solution, add glutaraldehyde solution with a concentration of 2.5% by volume, and fix for 2 hours at 4°C with...
Embodiment 3
[0056] The specific steps of the method for preparing Aspergillus niger transmission electron microscope ultrathin section samples with lead solution as vital staining agent:
[0057] (1) In vivo staining: Take a sterilized 250ml Erlenmeyer flask, add 100ml TAP liquid medium, transfer 1 mL of Aspergillus niger spore liquid into the medium with a pipette gun, and then transfer 100ul with a pipette gun at a concentration of 1000mg / L of sterile Pb(NO 3 ) 2 Solution enters culture medium (medium lead concentration 1000ug / L), seals with aseptic parafilm, shakes up;
[0058] (2) Cultivation: Place the Erlenmeyer flask after the inoculation in step (1) in a shaker at 180r / min in the dark at 28°C for 4 days;
[0059] (3) Pre-fixation: Take an appropriate amount of Aspergillus niger sample solution, add glutaraldehyde solution with a concentration of 2.5% by volume, and fix for 2 hours at 4°C without light;
[0060] (4) Rinse: wash the sample 4 times with phosphate buffer for 2 hou...
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