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110 Cards in this Set
- Front
- Back
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define bioluminescence
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the production of light by an organism, luciferin and oxygen are enzymatically catalysed by luciferase converting chemical energy into emitted light energy
squid, rail road worm, fire flies |
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The difference between bioluminescence and fluorescence
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fluorescence requires excitation light
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coomasie blue test
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meastures proliferation (cytotoxicity test) by total cell protein.
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wiscott-aldrich syndrome
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x-linked recesive disease presenting with eczema + thrombocytopenia and - IGm
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cyclosis (cytoplasmic streaming) examples
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movement of cytosol + chloroplasts in plant cells (elodea), actin microfilament based.
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MCA uses
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tumour marking
radio + chemotherapy (trastuzumab= herceptin) IHC |
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Apoptotic studies specimen.
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caenorhabditis elegans
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locomotion examples in the human body:
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cell crawling - mesenchymal cells
ameboid movement: macrophages growth cones: neurons |
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overtons rule
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inc lipid solubility inc rate of cell penetration
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GZMB
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granzyme B - protease used by T-ly in inducing apoptosis
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filopodia
lamellipodia |
f-actin microspikes project from the cell surface
flat dense actin meshwork betweek the microspikes |
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Cytoskeletal associated proteins:
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1- fascin to actin
2- talin anchors actin 3- myosin - vesicle transport 4- MDTA - microtubule actin links |
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Stress fibers
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bounds of microfilaments, cross linking proteins and myosin 2 motors.
change in cell locomotion- keep cell shape, stability |
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FAS-l (ligand)
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part of the "death receptor" family - a cell surface protein which binds to FAS-R to trigger apoptosis
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Colour spectrum
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blue: 450-500nm
yellow: 570-590nm red: 620-750nm |
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hayflicks limit
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20-80 generations of cells within a culture = shortening of telomeres reaches critical level.
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autofluorescing molecules
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haemoglobin
rhodopsin phycocyanin myoglobin |
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CDK
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cyclin dependant kinase
regulate the cell cycle at regular checkpoints |
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P21 / waf
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Inhibits CDK2 thus regulates the cell cycle and suppresses cell proliferation
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P53
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Tumour supressor protein, conserves the genome by initiating dna repair and preventing mutations.
It can INDUCE apoptosis |
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What is cell crawling?
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the specific rearrangement of microfilaments AND microtubules
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Ameboid movement?
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microfilament based rearrangement of cytoplasm and the formation of pseudopods
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Occurance of physiological apoptosis
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1 embryological development
2 elimination of cells with damaged dna 3 cellular homeostasis 4 involution of thymus 5 pos/neg selection of T-ly in thymus 6 barriers (sertoli cells) |
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trypan blue stains?
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Lysosomes blue in Hep-2 cells
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Increasing magnification
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dec - depth of field
dec - field of view dec - working distance |
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what is Rb protein?
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retinoblastoma protein
in a hypophosphorlyated state it binds to regulation sequences of genes that are necessary for proliferation. |
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role of cyclin B - CDK1 complex?
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stimulates mitosis at metaphase
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cyclin D - CDK4 complex?
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Rb protein phosphorylation = stops proliferation
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PCR
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polymerase chain reaction
denaturation: 94*C annealing: 54*C elongation: 72*C exponentially amplifies DNA invitro |
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Li-fraumeni syndrome
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P53 mutation
damaged dna replicates with no regulation.- early age cancers (sarcoma mainly) |
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hereditary retinoblastoma
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mutation in Rb1 gene - resulting in cancer of retinal cells
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xeroderma pigmentosum
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mutation in genes for excision repair of DNA
thus the inability to repair damage caused by UV light |
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FAP/C
Familial adenomatus polyposis coli |
mutations in APC, loss of DCC and other mutations result in polyps in the colon, can be malignant or bleed often.
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confocal fluorescence advantages over WA fluourescence
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more sensitive,
3D imaging signal to noise ratio better automated why better? confocal has excitation and emission pinhole filter and uses laser scanning |
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what does a flow cytometer do?
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measures the ammount of DNA in a cell as it undergoes mitosis, this is done by staining the dna as it is replicated
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Mitotic index
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measure of the proliferation status of cell population, ratio no of cells in mitosis and total no of cells
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stress fibers
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microfilaments bound by proteins and myosin 2 motors
contraction can take place, and SF are anchored by focal adhesions to the cell membrane |
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Mesenchymal movement
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ruffling membrane, filopodia, lamellipodia.
RM contains receptors and cell adhesion molecules |
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discovery of GFP
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aequoria victoria - green fluorescence under UV, aequorin protein gave blue chemiliuminescence
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HEP2 cells are?
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continuous cell line from laryngeal carcinoma
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DNA extraction- chemically!
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SDS and EDTA breaks down PLBL and chelates out metal ions of proteins
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DNA extraction- enzymatically!
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Trypsin, EDTA, Collagenase
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cultivation mediums?
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CADMEC
melanocyte GM Fibroblast GM Eagles mem (minimal essential medium) |
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Which molecules show photodynamic effect?
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Porphyrins, chlorophyll, aminolevulinic acid, eosin
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Clinical treatments using PDT?
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Melanoma
Acne esophageal cancer wet aged macular degeneration |
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Ionising radiation effects on dna
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point mutation
chromosomal breaks (ad, del, rearr) mitotic defects |
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A stable cell line has been...
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sub cultured (passaged)
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Types of apoptosis (4)
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normal - (ex/in pathways, A-bodies, singular, blebbing, PPS externalisation
Anoiki- anchorage dependant cells detach from the ECM which induces A Mitotic catastrophy- mitotic linked cell death, ch segregation or DNA damage Oncosis- Ischemia, mitochondiral swelling and cytoplasm vacuolisation. |
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microtubule construction
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heterodimers consisting of a-tubulin(+) and b-tubulin(-)
negative from MOC/centrosome, positive to cell membrane |
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Jodoanal B to test..
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cell viability via iodophor
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Types of cell fusion
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spontaneous
induced: chemicals, electrically, sendai virus |
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dissagregation
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process of deystroying proteins of junctions that interconnect cells.
EDTA, trypsin, collagenase |
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what is blebbing?
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cytokeratinisation fragmentation during apoptosis
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depth of field
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the area infront and behind the specimen that will be in acceptable focus
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Fluorescing labels
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phalloidin (red) - actin
FITC (green) - DNA for flow cytometry |
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Fluorescing probes
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non-covalent (native stains)
janus green, acridine orange, neutral red |
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coffilin
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actin binding protein which regulates the assembly and dissasembly of actin filaments
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gelsolin
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actin binding protien which regulates ass/diss of actin (but needs Calcium to induce its function)
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Integrin
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Is a receptor which binds the cell to the extracellular matrix
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Vinculin
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links integrin to the cellular actin cytoskeleton
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The difference in lenses between confocal and normal fluorescence microscopes
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confocal = excitation + emission pinholes. One area of the specimen can be selected = high resolution
normal = just a lens |
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epi-illumination
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light via the objective illumintes the specimen, it is then recollected by the same objective lens to the viewing piece
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cell fusion depends on?
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proteins of the cell membranes to mutually interact and join
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Apoptosis activatin factors
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TNFR, APOP1, Fas
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crenation
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osmotic responce to RBS in a hypertonic solution
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Eukaryotic DNA
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linear, histone associated, introns, exons
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main advantages of confocal microscopy?
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improved resolution- axial resolution of thicker specimens
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Extracellular apoptosis triggers?
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hormones, growth factors, cytockines, nitric oxide
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Positive apoptosis?
Negative apoptosis? |
binding and subsequent A
repression of A by a molecule is stopped |
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Agar diffusion principle?
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measures the effect of an antimicrobial agent against bacteria in a culture
endpoint- no colonies will grow where the concentration in the agar is at an effective level |
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SDS
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sodium dodecyl sulphate:
disrupts non-covalent bonds in proteins= denaturation |
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3R principles
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Russel + Birch 1959
"principles of humane experimental techniques" reduction, refinement, replacement |
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nonidet solution
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breaks down membranous structures
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cytotoxicity test principles in vitro
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seed, cover, add, test, decolour agar
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fluid mosaic model
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dynamic relationship between membrane proteins and phospholipid bilayer
singer and nicholson 1972 |
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FRAP
FRET BRET |
fluorescence recover after photobleaching
fluorescence resonance energy transfer (flurochrome excites other molecules around it) Bioluminescene resonance energy transfer (fire flies) |
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stain for endoplasmic reticulum
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DioC6 - green
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DNA stains
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dapi - blue
hoechst - blue ETbr - orange Acridine orange - green/ orange |
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taxol
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stabilises microtubules resulting in G2/M phase arrest
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actin stain
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phalloidin = red (phallotoxin)
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EBESCO
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Elton B Stephens COmpany
One department is a database for scientific journals |
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Define a gene
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a lenght of dna at a locus on a chromosome which expresses a certain phenotype
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microfilament inhibitors
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phalloidin, swinholide, latrunculin, cytochalasins
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Microfilament rearrangement in locomotion:
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lamellipodia
filopodia stress fibers focal adhesions |
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cellular mitotic checkpoints
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restriction CP
G1 CP G2 co metaphase is the final point before cytokinesis |
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intracellular parasites
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salmonella, listeria, shigella, mycobacterium,
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cytoskeletal defects
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duchennes muscular dystrophy - dystrophin mutation cells not attached to BL
amyotrophic lateral sclerosis (mnd/ lou gehrigs) - inclusions in axons, UL/LL mn death |
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intracellular movement:
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vesicle trafficking, cyclosis, cytokinesis, pigment movement, vesicle discharge, endo/exo cyctosis
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UV absorption wavelengths for
nucleic acid proteins |
dna- 260nm
proteins- 280nm |
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two photon microscopy
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simultaneous absortion of photons allows mag of thicker sections, higher density of photons in the (infrared spectrum), decreases background signal
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Mag / SE equations
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drawing mag = actual drawing (um)/ SE (um)
SE = FOV / tangent no of cells |
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disadvantages of phase contrast microscopy
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flat specimens
halo effect round objects = flash of light resolution limit |
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lgd
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lou gehrigs disease
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dmd
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duchennes muscular dystrophy
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isi
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institute for scientific information
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sci
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scientific citation index
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pbl
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peripheral blood lymphocytes
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ctl
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cytotoxic t- lymphocytes
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hla
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human leukocyte antigens
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lcl
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lymphoblast cell line
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ncbi
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national center for biotechnological information
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bcl-2
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B cell lymphoma 2
anti/pro apoptotic proteins |
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Differentiation mechanisms of stem cells?
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fusion with target tissue's cells
gene expression changes resulting in transformation into new target cell (terminally differentiated) |
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hNT- cells are what?
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human non-tumorigenic cells.
Derive from teratocarciomas- tamed in a lab and now have neuronal cell generating capacity. |
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OEG cells
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olfactory ensheathing glial cells
source of glial stem cells in the olfactory bulb |
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SHED
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'stem cells from human exfoliated decidous teeth'
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SCID
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'severe combined immuno deficiency syndrome'
Gene therapy can insert a gene into pt's BMSC and cure the dissorder. |
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FLIM
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fluorescence lifetime imaging microscopy-
the lifetime of the flurophore signal rather than intensity creates the image two photon microscopy |
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two photon microscopy
three reasons for deeper sectioning |
multiphoton absorption at one point
Photons in the IR spectrum = more penetration FLIM |
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PBS
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phosphate buffered saline
used in peridontal ligament fibroblast extraction |
