Microscope stage incubator

Imaging of live OCCs requires a microscope stage incubator. To perform live cell experiments start by filling the medium reservoir of the microscope stage incubation chamber with culture medium. SrrNote 19. 

Then transfer a culture on a Millicell-CM insert in the microscope stage incubator, close the chamber lid and turn on the CO2 supply. Allow culture to equilibrate and then choose a microscope field with several transfected cells using the conventional fluorescence microscopy mode of the confocal microscope. 

Perfuse the antiplatelet antagonist at the desirable concentration or vehicle (control) over surface-bound platelets, and incubate for 10 min. The extent of platelet activation can be further modulated by the presence of chemical agonists such as thrombin (0.02-2 U/ml) during the 10 min incubation. The microscope stage and flow chamber are maintained at 37 °C by an incubator heating module and incubator enclosure during the experiment. 

Incubate the cells on the MatTek dish in a humidifled, 37°C, 5% COj incubator or on the microscope stage in an environmental chamber with similar conditions for about 3 h and then start to observe with Olympus TIRF microscopy. 

Wipe all tissue culture incubators, hoods, tissue choppers, and microscope stages with 70% ethanol on a regular basis. It is advisable to have a dedicated tissue chopper that remains in a sterile tissue culture hood. Most contamination problems occur during the preparation of the slice cultures. Contaminants are generally of two types yeast and bacteria. Control bacteria by adding chloramphenicol... 

For ligand-gated ion channel receptor expression studies, stage V and VI oocytes are preferred (see earlier). Under a dissecting microscope, oocytes of this stage that are in good condition are sorted from the rest and then incubated at 16-18°C in MBM overnight. Incubation in hypertonic phosphate... 

Hens and quails eggs incubated 40 4h so that they are at stage 10-12. The hens eggs (hosts) should have been resting on their sides at least 30 min. Dissecting microscope. 

Already at its earliest stages the colorless potential hemocytoblast cells of the blastoderm contain all the enzymes that convert ALA to porphyrins at nonlimiting activities. This was shown by incubation of the blastoderm with ALA. Porphyrinogen intermediates accumulated within hours, some of which became autoxidized to porphyrins such as coproporphyrin and protoporphyrin. These porphyrins could be detected in cells with a fluorescence microscope. 

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