TLC Flashcards
What are the 2 types of electron microscope?
> Scanning.
> Transmission.
How does a Transmission electron microscope work?
> Fires electrons through a thin sample.
> Electrons are seen by the detector.
What is an electron lense?
An electromagnet used to focus electrons
How does a Scanning electron microscope work?
Detects electrons reflected of the samples surface.
What are the disadvantages of electron microscopy?
> Samples must be sliced extremely thin. > Sample prep requires the use of toxic chemicals (heavey metals). > Sample prep can alter tissues. > Expensive. > Requires specialist knowledge.
How is it possible to detect specific proteins using light microscopy?
> Linking antibodies to florescent labels.
> Linking antibodies to gold beads.
Cytology is the study of what?
Structure and function of cells.
Histology is the study of what?
Microscopic structure of tissues and organs.
Pathology is the study of what?
Abnormal structure and function.
What is the function of the cytoskeleton?
Controlling the shape and mobility of the cell.
What are 3 ways it is possible to see parts of cells?
> Cut the tissues into slices.
Remove individual cells.
Use specialised imaging techniques (staining).
How do you prepare slides (bright field microscope) ?
> Fixation (preserves). > Embedding in wax (makes more rigid). > Cut into sections. > Mounting. > Staining. > Covering.
What colour do Haematoxylin and Eosin stain?
> Haematoxylin - DNA blue
> Eosin - proteins pink
What are the 5 main parts of a light microscope?
> Eyepiece. > Objective. > Sample. > Condensor. > Collector lense.
What are some advantages of bright field microscopy?
> Sample prep simple.
Clear images of overall tissue morphology.
Quick imaging.
What are some disadvantages of bright field microscopy?
> Images low contrast. > low sensitivity. > Samples must be stained. > Can not image living cells! > Colour resolution poor.
What is phase contrast microscopy?
Microscope designed by Fritz Zernike where 2 rings are inserted into the light path; one before the sample, one in the objective.
What are some advantages of phase contrast microscopy?
> Quick to set up.
> Can be done on living samples.
What are some disadvantages of Phase contrast microscopy?
> Produces poor spatial resolution.
“halo” produced hiding detail.
Not sharp.
What is Differential interferance contrast microscopy?
Light microscopy wher components are added; 2 polorizers and 2 prizms.
What are advantages of Differential interference contrast microscopy?
> Very good resolution.
Sharp.
Can be done on live samples.
What are disadvantages of Differential interference contrast microscopy?
> Complex to carry out.
Sensitive configurations.
Can not be done with samples in plastic dishes.
How other than by using stains can parts of a cell be labelled?
Using fluorophores.
How does Fluorescence work?
> Bright light shone through filter.
Light reflected on to sample by a dichroic mirror.
Light given off collected through filter.
What is “pseudocolouring”?
Adding a false colour to make image clearer.
What are the advantages of Fluorescence microscopy?
> Increased sensitivity.
Used to distinguish 4 or more different stains.
Used for live cells.
Non-toxic.
What are the disadvantages of Fluorescence microscopy?
Dyes bleach as light is shone on them.
What is Immunostaining?
Binding a coloured antibody to a selected protein.
What is the Cytoskeleton made of?
> Actin - filaments.
> Tubulin - microtubules.
How are proteins made?
> Proteins destined to live inside the cell are made by ribosomes.
Proteins destined to live within membranes or be transported out of the cell follow a more complex path.
How are intracellular membranes distinguished between under microscope?
> Proteins they contain.
> How they move.
How are lysosomes recognised?
Using a red stain activated under acidic conditions.
What are the main components of a (eukaryotic) cell?
> Plasma Membrane. > Cytoskeleton. > Nucleus. > Mitochondria. > Endoplasmic reticulum. > Golgi. > Secretory vesicles. > Endosomes. > Lysosomes.
What is hyperplasia?
Cell division.
What is hypertrophy?
Increase in cell size.
How are exact copies of DNA made in cells?
Semi-conservative replication.
When does mitosis occur normally?
During early embryogenesis.
When does mitosis occur pathologically?
> Benign hyperplasia of the prostate.
> Development of mammary tumours.
What happens during hypertrophy?
Extensive biosynthesis.
When does hypertrophy occur naturally?
In structures such as endochondral bone formation.
When does hypertrophy occur pathologically?
> Obesity - adipocytes enlarge.
> Hypertrophic heart disease - cardiomyocytes enlarge.
What are the 3 inter-dependant cell cycles?
CYTOPLASMIC cycle.
CHROMOSOME cycle.
CENTROSOME cycle.
What happens during the cytoplasmic cycle?
Growth and biosynthesis fuels:
> Chromosome and centrosome cycles.
> Organelle replication.
> Seperation of daughter cells.
What happens during the chromosome cycle (mitosis) ?
> DNA replication.
> Segragation.
When does the centrosome cycle happen and what happens?
> Prior to mitosis.
> Formation of the mitotic spindle fibres.
Whats the difference between a centrosome and centriol?
Centrosome consists of 2 centrioles.
What happens during cell cycle arrest?
Halt in progression through cell cycle, hypertrophy still occurs.
When during the cell cycle do the checkpoints take place?
> Between G1 and S phase (restriction).
Between G2 and prophase.
During metaphase of mitosis (spindle)
What’s the point of a checkpoint 1 (restriction)?
> Ensuring DNA is not damaged.
> Ensuring cell is large enough.
What’s the point of checkpoint 2?
> Ensuring DNA is not damaged.
> Ensuring DNA replication is complete.
What’s the point of checkpoint 3?
> Ensuring mitotic spindle is assembled.
> Ensuring chromosomes are properly aligned?
When does the double check occur?
During checkpoint 2, it ensures that there is no error in duplication of chromosomes.
What happens during the G1 phase?
Normal cell activity (biosynthesis) production of organelles.
What happens during the S phase and how long is it?
> Cell content doubles (DNA + cytoplasm).
Chromosome forms 2 sister chromatids.
S phase is 3-6 hours.
What happens during the G2 phase?
> Normal cell activity (biosynthesis).
2-4 hour delay.
Cell cycle checkpoint.
What happen if cells become locked in G1 phase?
HYPERTROPHY
What happens if cells have unrestricted acces to the mitotic phase?
HYPERPLASIA
Which is the longest phase?
G1
What happens during the M phase and how long is it?
> Protein synthesis ceases.
Chromosomes condense.
1 hour.
What sort of cells spend time in a resting state G0 phase?
Cells dividing less frequently than once a day.
What happens in the G0 phase?
Cells either become QUIESCENT or undergo HYPERTROPHY.
What part of the cell cycle can be transient or permanent?
G0 phase
how do cells reduce the length of their cycle e.g. during the rapid growth of embryonic cells?
Through virtual ELIMINATION of G1 and G2.
How can cells in each steps of the cell cycle be quantified?
> Estimated by microscopy.
Labelled with radiolabelled niwcleotides.
Using flow cytometry.
How is control of the cell cycle maintained?
Decision points and Checkpoints.
What are features of the cell cycle?
> Time limited (has a clock)
Invariable order.
Phases are irreversible.
Robust - works even after malfunction.
What do cyclins and CDK’s do?
Regulate cell cycle activity.
What happens when CDK’s are active?
PHOSPHORYLATION - progression of the cell cycle.
What happens when CDK’s are inactive?
No phosphorylation - cell cycle is halted.
What must happen in order for an EXPLOSIVE RISE in CDK binding and formation of the ACTIVE COMPLEX?
Cyclin concentration must reach a THRESHOLD