Cell Biology Staining And Counting

Front 1

For microbiology, cell culture, and many applications that require use of suspensions of cells...what is necessary?

Back 1

to determine cell concentration

Front 2

A device used for determining the number of cells per unit volume of a suspension is called a _____.

Back 2

counting chamber

Front 3

The most widely used type of chamber is called a _______

Back 3

hemocytometer

Front 4

the most widely used chamber is called a hemocytometer, since it was originally designed for ____

Back 4

performing blood cell counts

Front 5

to prepare the counting chamber the mirror like polished surface is carefully cleaned with _____. the ______ is also cleaned.

Back 5

lens paper

cover slip

Front 6

coverslips for counting chambers are specially made and are thicker than those for conventional microscopy..why?

Back 6

since they must be heavy enough to overcome the surface tension of a drop of liquid

Front 7

when is the coverslip placed over the counting chamber?

Back 7

prior(before) to putting on the cell suspension

Front 8

the suspension is introduced onto one of the V SHAPED WELLS with a _____

Back 8

Pasteur or other type of pipette

Front 9

the area under the coverslip fills by ______

Back 9

capillary action

Front 10

enough liquid should be introduced so the the ______ is just covered

Back 10

mirrored surface

Front 11

the charged counting chamber is then placed on the microscope stage and the _____ is brought into focus at low power

Back 11

counting grid

Front 12

why is it essential to be extremely careful with high power objectives?

Back 12

sine the counting chamber is much thicker that a conventional slide

the chamber or an objective lens may be damaged if the use it not careful

Front 13

one entire grid on standard hermacytometers with NEUBAUER RULINGS can be seen at ___

the main divisions separate the grid into ___ large squares

each square has a surface area of _____, and the depth of the chamber is ___

thus the entire counting grid lies under a volume of _____

Back 13

40X (4X objective)

9 large squares

each square : one square mm

depth of chamber: 0.1mm

volume : 0.9mm cubed

Front 14

how dilute should the suspensions be?

Back 14

dilute enough so the cells or other particles do not overlap each other on the grid, and should be uniformly distributed

Front 15

for cells that overlap a ruling count a cell as "in" if it overlaps the ______ ruling, and "out" if it overlaps the ______ruling

Back 15

in top right
out bottom left

Front 16

in many cases, it is necessary not only to determine how many cells there are, but also to determine the _____

Back 16

number of live cells

Front 17

The ratio of live cells to total cells is __________

Back 17

the cell viability

Front 18

ratio of live cells to total cells (cell viability) is an important parameter in _____&______.

Back 18

cell cytotoxicity test and cell culture

Front 19

the easiest and most widely used technique for determining cell viability (not the most accurate though) is measure cell viability is ________

Back 19

trypan blue staining

Front 20

what is trypan blue?

Back 20

a dye that is used to determine the cell viability

Front 21

when using trypan blue as a dye to determine the viability of the cells living cells _______, whereas dead cells will ____. the blue stain is easily visible, and cells can be counted using a light microscope.

Back 21

exclude the dye
dead cells will take up the blue dye

Front 22

goal for this weeks lab:

we will count the number of cells from ____, and add ______on a petri dish

we aim to grow these cells till we use them to estimate total cellular protein for our next cell bio lab.

Back 22

T25 flasks

10 to the 5 cells per ml

Front 23

what are your 3 materials?

Back 23

media pre warmed to 37 degrees Celsius
T25 culture of macrophage cells from previous week
0.4% trypan blue

Front 24

what is your equipment (8)?

Back 24

1 sterile gloves, and lab coat
2. water bath to appropriate temp
3. microbiological safety cabinet at appropriate containment level
4.CO2 incubator
5.35mm cell culture dish
6.marker pen
7.pipettes
8.petri dishes

Front 25

how do you calculate percent of viable cells?

Back 25

# of unstained cells / # of total cells times 100