Mitosis Flashcards
importance of growing cells in culture
allows complete control of growth conditions
why are cells from frogs used
cold blooded
continue to divide actively at RTP
free from potential hazards associated with human cell lines
culture medium
complex mixture of slats nutrients vitamins other factors - promote growth and cell division
pH indicator
puce ten purple - alkaline
clear then orange - acid
vinablastin
used as part of chemo
inhibit cell division
toxic to all cells
in chemo - preferentially targets rapidly dividing cells
process of applying the inhibitor
remove cover slips from petri dish
pick up coverslip using duck-billed forceps (close to edge so don’t scrape and damage cells)
transfer to slot in Columbia jar - containing drug - with the cells facing the spot in one side of jar
metaphase
chromosomes fully condensed
nuclear envelope break down - form vesicles that reassemble at end of mitosis
chromosomes lie on the metaphase plate at centre of mitotic spindle
microtubules attach to centromere via kinetochore
replicated dna
chromosome not linear
exist as 2 chromatids
identical DNA
joined by centromeres
staining procedure
dry forceps - avoid contamination
take coverslip
wash in phosphate buffered saline by dipping under surface 5 times
fix cells - dip into acid alcohol and hold for 5secs
rinse by dipping 5 times in distilled water
place 1 drop of neutral red stain in centre of coverslip - lower slide on it
examine under microscope - identified by dense staining chromosomes at 40x mag
prophase
individual chromosomes condense and take up stain
chromosomes exist as chromatids
nucler envelope breaks down
interphase staining
nucleus stains weakly except for nucleoli
anaphase
sister chromatids separate
microtubules attach to kinetochores on centromere and pull 1 chromatid to each pole
telophase
2 separate groups of chromosomes form nuclei of daughter cells
chromosomes de-condense
nuclear envelope reforms
end of telophase have cytokinesis - see cleavage furrow
counting cells
scan systematically
