microscope Flashcards
what is the standard protocol for using the microscope?
Place and adjust position of slide on stage
Adjust eyepieces to suit your eyes
Use of different objectives
Adjustment of brightness
Adjustment of the aperture of the condenser iris diaphragm
Set focus of condenser lens
Recording the position of an object on a slide.
working distance
The distance between the edge of the objective lens and the specimen surface
objective lens 4x
18.5 mm working distance
objective lens 10x
10.6 mm working distance
objective lens 40x
0.6mm working distance
objective lens 100x
0.13 mm working distance
Resolution
refers to the ability of the instrument to distinguish between two points. The clarity of the image. resolution is defined as the minimum distance 2 points can be separated and still be distinguished as 2 separate points.
in-vitro cell culturing of HeLa cells
key is to maintain sterility.
incubator: temp of 37, 5% CO2 and humidity.
cell culture media: growth factors (FCS), amino acids (L-Glutamine), antibiotics etc.
% confluency
The estimated % surface area the growing cells have taken up
Fluorochromes
are fluorescent dyes that attach to visible or sub-visible structures, highly specific
DAPI (4’,6-Diamidino-2-phenylindole)
Forms fluorescent complexes with AT-rich sequences of double-stranded DNA
Acridine Orange
combines with DNA and RNA: DNA+AO = yellow/green; RNA+AO = red/orange
Rhodamine 123
a mitochondria-specific dye
Immunofluorescence
use of antibodies to which a fluorescent marker has been attached.
antibodies
are molecules that recognize and bind selectively to specific target molecules in the cell
