Techniques (Bio 311, Test 1)
cDNA Cloning
cDNA Cloning starts with mRNA
ex. of Nomarski/DIC
single cell electrophysiologist, Amoeba and C. elegans embryo
Atomic Force Microscopy
-AFM - No glass lenses - Fine tip - E.g. nuclear pores
Apoptosis/Necrosis/Annexin V/Propidium iodide
-Cell death - Apoptosis: programmed cell death, Anexis V binds to phosphatidylserine - Brazilian Wasp MP1 relies on redistribution of phosphatidylethanolamine and phosphatidylserine in cancer cells -kills cancer cells but not normal cells - Necrosis: pathological cell death ("Ricin": targets protein synthesis) - Normal = inner leaflet, apoptosis = inner leaflet
FRAP
-Fluorescence recovery after photobleaching - Many ways -Originally, the FRAP technique was intended for use as a means to characterize the mobility of individual lipid molecules within a cell membrane - Photo destroying the GFP, and then watching the repopulation into the bleached area can reveal information about protein interaction partners, organelle continuity and protein trafficking - Can reveal exchange rates or membrane fluidity - Cannot monitor the same proteins as FRET -
Vital fluorescent dyes
-Fluorescent dyes to analyze changes in cell behavior or physiology in living cells - 1980s -Problem: dye → dye-am (am = acetoxymethyl) - Examples: Mitochondria activity: Rhodamine 123, JC-1, Live/Dead assay: calcein-AM, propidium iodide, used when looking at cancer cells, Ca2+ in living cells: Fluo3-AM and others, micro spectrofluorometry
FRET biosensor
-Fluorescent protein biosensors have found widespread utility in reporting on a diverse array of intracellular processes - Spectral imaging has been very useful for the examination of fluorescent protein biosensors to determine the presence or absence of FRET in response to a biological stimulus - Thus, by creatively fusing pairs of fluorescent proteins to biopolymers that perform critical functions involved in various aspects of physiological signaling or other biological activities, a number of investigators have developed a host of new molecular probes that are useful for optical live-cell imaging of important metabolic and signaling processes
fluorescence microscopy
- 1970s - Elements: fluorochromes, many fluorescent techniques - Fluorochromes: fluorescent dyes, excitation (wavelength = 485 nm) and emission (535 nm)
Biological scaffolds
- A key requirement for tissue engineered constructs - 3D - Some biodegradable and others not - ELAD, human bladders, cartilage (carticel) - Hepatoma cell line is 50% cyt P450 → hepatoma cells cause cancer
Pulse chase
- A method for examining a cellular process occurring over time by successively exposing the cells to a labeled compound (pulse) and then to the same compound in an unlabeled form (chase) - This method is useful for determining the activity of certain cells over a prolonged period of time
GFP, YFP, CFP as reporter molecules
-Green fluorescent protein - Jellyfish - Recorder molecule of expression - E.g. Application: if gene expressed, the cell is green - Roger Tsien: 2008 Nobel Prize for GFP Discovery - Many fluorescent proteins means multiple labels - Can reveal protein expression in whole organisms as well as cells - Ex of use: Transfected Androgen Receptor in PC-3 prostate cancer cells
High voltage electron microscopy
-HVEM - Theoretical limit of resolution is better because accelerating voltage is 1000kv, not 100kv as in typical,TEMs but best used for imaging thick sections
2D gel electrophoresis
-Higher resolution than the previous ones - 1D = IEF (Isoelectric focusing) - 2D = SDS
Autoradiography (extended history)
-1970s - Radioisotopes - E.g. 3H-thymidine: labels DNA, 3H-leucine: labels proteins - Identifying Dividing Cells in Breast Tumors using 3H-thymidine - Can track cell processes over time
EGTA/Protease
- Calcium chelator: decreases the activity of a calcium dependent cell adhesion molecule - Trypsin: protease: cleaves proteins such as ECM proteins - Application: cell therapy such as pancreatic islets transplantation, research - Used in combination to facilitate tissue dissociation
PCR
- Amplifies DNA - Key enzyme: Taq polymerase (extremophile) - DNA polymerase is heat resistant
Bioprinting/3D printing
- Bioprinting scaffolds (biodegradable) part of tissue engineering (stem cells) - Can print tissue by swapping ink jet printer for suspension of cells -Rapid prototype: group of techniques used to quickly fabricate a scale model of a physical; part or assemble using 3D computer aided design -1980s ink jet printer
Southern blots
- Blotting technique - Designed to detect genes - Qualitative: molecular weight - Quantitative: abundance - Polymorphic markers = you are almost unique (e.g. Variable Number Tandem Repeats -Useful in Prenatal Diagnosis, Medicine and Crime), bone marrow transplant
Protein A immunoprecipitation
- Antibodies - Protein A (S. aureus) - Binds to all types of antibodies - Fc region - Process can be used to isolate and concentrate a particular protein from a sample containing thousands of different proteins - Precipitating a protein antigen out of solution using an antibody that specifically binds to that particular protein
Western blots
- Antibodies to identify select proteins of interest - Qualitative: molecular weight - Quantitative: relative abundance - Tracking Ligand- receptor interactions in living cells, gel electrophoresis, blot: transfer paper - Ex. W/ photoreactive amino acids - Built on a technique that involves transferring, also known as blotting, proteins separated by electrophoresis from the gel to a membrane where they can be visualized specifically
Tissue engineering
- Artificial blood vessels: the "Holy Grail" of tissue engineering - Anthony Atala's Group: Clinical implants of tissue engineered neovaginal organs using biodegradable scaffolds, autologous muscle and epithelial cells- Mayer-Rokitansky-Kuster-Hauser syndrome - Can be used in 3D screening to identify drugs that might target pancreatic cancer or secreted biomarkers - Used for testing and optimizing thermal ablation medical devices - Subdiscipline of regenerative medicine, use of cells to generate tissue, "organoids," 3D - Bad start with "vacanti mouse" by putting a human ear on a mouse
Band/gel shift assays
- Band assays are useful to look at DNA binding proteins - Isolate protein and look at gene of interest within proteome
Mutations
- Can be caused by ultraviolet light, chemicals, etc - Absolutely critical in molecular genetics
spinning disk microscopy
- Can reveal changes in endoplasmic reticulum in living HeLa cells - Many spots simultaneously - Virtues: faster, less laser intensity, less photobleaching and heat, living cells and analyzing dynamic activities in milliseconds - The sample is illuminated and light detected simultaneously at multiple points
phase contrast microscope
- Cell biologists (most common for cell cultures) - Living cells, does not use dyes (flourochromes or colorimetric) - Manipulate the light path to increase contrast - Converts phase shifts in light passing through a transparent specimen to brightness changes in the image - Phase shifts themselves are invisible, but become visible when shown as brightness variations
Transfection delivery methods
- Chemical transfection is a popular technique due to the ease, cost, and wide variety of transfection reagents available for purchase - Transient transfection is commonly used for short term expression of a desired gene that lasts a few days - Research applications include research of gene expression, gene silencing studies and analysis of recombinant proteins - Electroporation and cell injection (e.g. gene guns) are two classical examples of physical transfection methods that are able to accomplish delivery of nucleic acids, but are harsh on cells due to disruption of the cellular membrane and often results in cell death - It is an appropriate option for cells that have been traditionally difficult to transfect
Decellularization
- Decellularization is the process used in biomedical engineering to isolate the extracellular matrix (ECM) of a tissue from its inhabiting cells, leaving an ECM scaffold of the original tissue, which can be used in artificial organ and tissue regeneration
sources of cancer cells
- Derived from a tumor,Transfected with oncogenes - Chemicals that mutate cells - Spontaneous mutation (lost a tumor suppressor gene)
Freeze fracture
- Designed to look like membrane topology - Metal replica - A specimen is frozen rapidly and cracked on a plane through the tissue, this fracture occurs along weak portions of the tissue such as membranes or surfaces of organelles
Complementation analysis
- Determines if different recessive mutations are in the same gene or different genes - Whether a mating or other introduction of genetic material can complement a mutation and produce a wild-type phenotype -Stringency tests - Genetic/mutation analysis
Densitometer
- Device that measures the degree of darkness (the optical density) of a photographic or semitransparent material or of a reflecting surface - Light source aimed at a photoelectric cel - Converts bands to bars
Nomarski/DIC
- Differential interference contrast microscopy - Cell biologists - 3D image - Important for single cell electrophysiology and patch clamping (injection) - Used to enhance the contrast in unstained, transparent samples - Living cells - DIC works by separating a polarized light source into two orthogonally polarized mutually coherent parts which are spatially displaced (sheared) at the sample plane, and recombined before observation
Dynabeads/Speedbeads etc.
- Dynabeads are frequently used for cell isolation (cell separation) - Cell-types often of interest to purify may be specific leukocytes, such as CD4+ T cells, stem cells,[7] or circulating tumor cells (CTCs) - Spherical polymer particles with a uniform size and a consistent, defined surface for the adsorption or coupling of various bioreactive molecules or cells
Knockout /knockin mice and cells
- Edit genes out - Used to study gene function, usually by investigating the effect of gene loss
Veridex CellSearch System
- Enumerate circulating tumor cells (CTCs),- Method for detecting low number ( 1 in a billion), involves magnetically separating CTCs from other cells - Circulating Tumor Cells (FDA approved) are a sign of metastasis
Laser capture microdissection microscopy
- Ethylene: vinyl acetate film - Melted with a laser - Remove → analyze a cell of interest - One or two cells can be removed from a heterogeneous tissue section or group of cells for molecular analysis
FISH
- Fluorescence In Situ Hybridization,- For detecting mRNA (e.g. detect specific mRNA in dendrites) - Designed to detect gene sequences or messenger RNA in situ (in place) inside cells - Uses fluorescent probes that bind to only those parts of a nucleic acid sequence with a high degree of sequence complementarity - Used in genetic counseling, detecting HPV in HeLa cells, medicine, species identification
Ultrastructural autoradiography
- Ex: 131-I labeling of the thyroid and 131-I α−bungarotoxin labeling ACH receptors - Detection method in which X-ray or photographic film is exposed to emissions from radioisotopes on TLC plates to produce an image on the film
cryostat
- Fast method - Frozen sections - Adv: fast - Dis: poor cell architecture
Equilibrium Density (Rate Zonal) centrifugation
- Ficoll used: it separates our mononuclear cells from plasma in the blood, this process is needed to separate cells and isolate organelles - Rough settles above smooth → separated by densities - The tube is first filled with different concentrations of sucrose or another solute establishing layers with different densities and viscosities, forming a density gradient, within which the particles to be separated are added - The larger particles will be able to travel to the bottom layer because they are more massive
Plastic thin sectioning
- Fix the cells using glutaraldehyde of OsO4 - Dehydration,- Infiltrate w/ plastic - Cut using diamond knife →ultramicrotome - Sections: suspend on a copper grid - Stains only heavy metals e.g. uranium for cytoplasm and Pb for membranes
Guava
- Flow cytometry - Doesn't separate cells but can enumerate (count cells) - Quantitative - Flow cytometry but using microcapillary tubing = small samples
Two photon microscopy
- Fluorescence - Abbe's equation limited - Deeper tissue penetration, e.g. brain tissue in vivo, will not create adverse phototoxicity when excited by your microscope system
FACS/Flow cytometry
- Fluorescent activated cell sorting - Cell separation technique - Needs a fluorescent probe - Not used directly for protein purification - Fluorescent label is cell specific - FACS machine: technology that is used to analyse the physical and chemical characteristics of particles in a fluid as it passes through at least one laser
FRET
- Forster resonance energy transfer - Proximity of two molecules - Biosensors: proteins tagged by two fluorescent proteins that can reveal change in cell physiology - E.g. Calmodulin: binds to Calcium → calmodulin changes shape → FRET biosensor → finishes green - A mechanism describing energy transfer between two light-sensitive molecules
Northern blots
- Gene transcription → mRNAs - To study gene expression by detection of RNA (or isolated mRNA) in a sample
Genomic Cloning
- Genomics: Controlling gene expression (Knockout/knockin mice) - Critical to "biologic" = humulin - Starts with all of the genomic DNA
cDNA microarrays
- Hybridization technique - Needs E.coli colonies, nitrocellulose filters, and autoradiography - Can analyze mRNA transcript - Adv: 1000s of mRNAs at once on "gene chip" i.e. insulin - Used for gene cluster analysis: patterns of similar expression
Fluorescence immunocytochemistry
- Identify the location of a particular protein in a cell, reveals cell polarity - Uses monoclonal or polyclonal antibodies, can use two different probes to get double label - Terms: Fluorochrome, Antigen (substance capable of eliciting an immune response → protein to be tracked), Epitope (the part of an antigen to which the antibody binds) - Direct: antibody with fluorophore attaches to antigen - Indirect: primary antibody is applied and then a second antibody with a fluorophore
darkfield microscopy
- Imaging of bacteria - Dark background - Light path: 1. light enters the microscope 2. a specially sized disc blocks some light from the light source, leaving an outer ring of illumination 3. condenser lens focuses the light towards the sample 4. the scattered light enters the objective lens, while the directly transmitted light simply misses the lens and is not collected due to a direct-illumination block 5. only the scattered light goes on to produce the image, while the directly transmitted light is omitted
Transfection
- Inserting intact genes E.g. GFP linked to gene of interest
Isoelectric focusing
- Isoelectric point: a point at which in a pH field the protein is electrically neutral and doesn't migrate - First dimension - Ex: blood serum (about 40+ bands) - Bands are very condensed - IEF
TIRF
- Looks at the edge of the cell - Other portion missing so it needs overlap with fluorescent confocal technique - High resolution allows for observation of membrane-associated processes
SELDI-TOF/MALDI-TOF
- Matrix-assisted laser desorption /ionization- Time of flight - Key parts: laser, "protein chips," TDF detection - Protocol: Use protein chips, Machine - Adv: small sample (1 microliter), resolution (1 amino acid), - Alzheimer's Disease ex: Generate an antibody (Beta-40 = normal, Beta-42 = Alzheimer's), MALDI-TOF peaks in unaffected shows largest peak at 40 with affected showing the largest peak at 42 - Uses a laser energy absorbing matrix to create ions from large molecules with minimal fragmentation (high resolution, size of one amino acid viewed clearly)
Microporous membranes
- Microporous membranes -When MDCK cells are grown on microporous membranes as they illustrate both the phenotype and function of its native cell type which is the Kidney -Cell culture growth substrates include: cell culture plates/dishes, beads, roller bottles, tubes, feeder layers for HESCs, and microporous membranes -SUMMARY: used as a filter in tissue engineering -Cultures can grow on them -May behave differently on this than any other substrate
Velocity sedimentation
- No dyes - Rely on differences in physical size → sedimentation rates - New microfluidic devices being developed - Cell separation technique
MRI microscopy
- Non-invasive/destructive - Spatial and chemical information - Resolution about 29 microns
ELISA
- Not a microscope technique, e.g. Fl. immunocyte = qualitative and ELISA = quantitative - Plate-based assay technique designed for detecting and quantifying substances such as peptides, proteins, antibodies and hormones - The antigen is detected either directly (labeled primary antibody) or indirectly (labeled secondary antibody) - The key step, immobilization of the antigen of interest, can be accomplished by direct adsorption to the assay plate or indirectly via a capture antibody that has been attached to the plate -Sandwich: most powerful ELISA assay format, the analyte to be measured is bound between two primary antibodies: the capture antibody and the detection antibody, best be used for the analysis of complex samples due to its high specificity and sensitivity
CRISPR/Cas9
- Research team used CRISPR to replace defects in heart tissue of Duchenne Muscular Dystrophy patients → Tissue engineered heart tissue now contracts better - Editing Genes - Adding/removing genes or single bases → CRISPR-Cas9 (62 pig retroviruses removed - on the way to xenograft transplantation? (animal to human) - Can be used to either delete single base pairs generating a frameshift mutation or repair or replace a targeted gene - 2018 - CRISPR-Cas9 babies born in China
Super Resolution Microscopy (3 types)
- Not diffraction limited (not limited by Abbe) - Three types: structured illumination microscopy (SIM), stochastic optical reconstruction microscopy (STORM)/photoactivation localization microscopy (PALM), and stimulated emission depletion microscopy (STED)
Yeast
- One tool that is often used in generating temperature sensitive mutant yeast - The "permissive" temperature for these yeast is most closely matched to 23 degrees Celsius
laser-scanning confocal microscopy
- Point scanning - Bright image - Seconds to generate - Fluorescent dyes used photobleach specimen, once they fluoresce they can't again - A single point of laser light is used to illuminate a sample. In this technique, laser radiation undergoes point by point scanning in a raster pattern. A photomultiplier tube is used to detect the signal sequentially from each point until a complete image is produced. - Trade-off between speed and resolution
DNA gel electrophoresis
- Polyacrylamide: small DNA fragments - Agarose: larger DNA molecules - Higher molecular weight at top and lower at bottom - Can tell an apoptotic cell population v. necrotic cell through DNA cleavage patterns
Polyclonal/monoclonal antibodies
- Polyclonal: inject antigen, B cells bind to the antigen, limited by protein kinase, bind to multiple epitopes and are usually made by several different plasma cell (antibody secreting immune cell) lineages - Monoclonal: top selling biologics, made by identical immune cells that are all clones of a unique parent cell, preferred because it only requires that one will have to bond to a specific site, can have monovalent affinity (they bind to the same epitope)
Radioactive tagging
- Process in which a regular isotope of an element is replaced by a radioactive isotope of the same element in order to track the presence and concentration of that element in a biological sample - This has applications in fields such as of biology by allowing biologists to track biological processes, and in medicine, by allowing doctors to track movement of particles throughout the body - Using imaging devices, doctors can then see things such as blood clots and tumors through radioactive tagging - Another application of radioactive tagging is following a biological process happening in vivo, such as cellular respiration
DEAE and CM Ion exchange chromatography
- Process that separates ions and polar molecules based on their affinity to the ion exchanger - Used directly for protein purification - Works on almost any kind of charged molecule—including large proteins, small nucleotides, and amino acids - The two types of ion chromatography are anion-exchange and cation-exchange
SDS gel electrophoresis
- Proteins are pretreated - Detergent - Used to solubilize membranes - Used directly for protein purification - SDS molecule: negatively charged, linear protein - Uses UREA (to break H bonds) and Beta-mercaptoethanol - Resolution increases, polymeric protein → monomeric components
Restriction nucleases
- RFLP's Restriction Fragment Length Polymorphisms -Used in genetic identity (finger print) and forensic medicine -Restriction Enzymes: cleave DNA at specified sites -Were originally isolated from bacteria -Palindromes are often discussed in reference to restriction enzymes (nucleases) -Most restriction enzymes recognize palindromic sites - Made it possible to cleave nucleotides into smaller, manageable pieces
Panning
- Selective surfaces: relying on a specific molecule and cell surface properly to achieve cell separation - An affinity selection technique which selects for peptides that bind to a given target - Cell separation technique - Conjugating the phage library to the desired target
Affinity Chromatography
- Separate proteins based on affinity - Used directly for protein purification - Ligand interaction - Antibody affinity → antibodies recognize proteins, purify antibody using antigens - Method of separating biochemical mixture based on a highly specific interaction between antigen and antibody, enzyme and substrate, receptor and ligand, or protein and nucleic acid
Gel filtration
- Separates proteins based on physical size and path length through a matrix - Beads: porous - Bigger protein comes out faster - Column chromatography - Used directly for protein purification
Single cell RNA
- Seq: track the transcriptome over time in living cells - An improvement over cDNA microarrays and can identify individual cells at select points in time based on their unique RNA signatures
Molecular beacons
- Similar to FISH - Fluorochrome + quencher - Can hybridize in situ in cells - Better than FISH - Probe's fluorescence is quenched (not fluorescent) unless bound to target RNA or DNA strand
Ultrastructural immunocytochemistry
- Similar to Fluorescence immunocytochemistry except it uses gold particles (nm), gold is used in ultrastructural immunocytochemistry instead of fluorochromes due to gold's high electron density which creates dark contrast spots by increasing electron scatter
SNPs
- Single Nucleotide Polymorphisms - Single Base Pair Site Where variation is found in at least 1% of population - Associated w/ personalized medicine-SNPs have been used recently to predict autism in babies and children - Its promise for personalized medicine appears strong as well - Common, "not mutations"
Temperature sensitive mutants
- Single point mutation give rise to proteins that are unstable and non-functional at slightly elevated temperature -Non Permissive Temperature - won't grow and proteins denature -Permissive temperature - proteins won't denature
Trituration/Sonication
- Sonication targets cell membranes, not for organelle isolation - Trituration for mechanical shearing - Apart of third step in cell separation
bright field/compound microscopy
- The condenser usually contains an aperture diaphragm to control and focus light on the specimen; light passes through the specimen and then is collected by an objective lens - Can be used by pathologists, histochemists, and cell biologists - Protocol: 1. isolate fixed tissue, cells 2. fixative i.e. formaldehyde cross links proteins 3. dehydration: remove water and replace it with ethanol 4. infiltration: replace ethanol with hot wax "block" of tissue: cut sections, microtome accomplishes this - This technique can be used to view fixed specimens or live cells - Adv: Superior tissue architecture - Dis: Low contrast, intense lighting can damage the specimen, SLOW - Uses colorimetric dye
Dialysis
- The process of separating molecules in solution by the difference in their rates of diffusion through a semipermeable membrane, such as dialysis tubing - The most common application of dialysis is for the removal of unwanted small molecules such as salts, reducing agents, or dyes from larger macromolecules such as proteins, DNA, or polysaccharides
Native gel electrophoresis
- Unaltered - Uses a gel typically made out of acrylamide - Proteins in the native state - Analysis: function, amino acid sequence, poor resolution - Power supply from (-) to (+), smallest travel furthest
Photoreactive amino acids,
- Upon exposure to ultraviolet light, they are activated and covalently bind to interacting proteins - In vivo or in vitro - They are ultraviolet light (UV)-activated to covalently cross link proteins within protein-protein interaction domains in their native in-vivo environment
Differential centrifugation
- Used to separate organelles and other subcellular particles on the basis of sedimentation rate] - In a typical case where differential centrifugation is used to analyze cell-biological phenomena (e.g. organelle distribution), a tissue sample is first lysed to break the cell membranes and release the organelles and cytosol - The lysate is then subjected to repeated centrifugations, where particles that sediment sufficiently quickly at a given centrifugation force for a given time form a compact "pellet" at the bottom of the centrifugation tube - Low speed: cells homogeneous, medium speed: pellet contains whole cell nuclei cytoskeletons, fast speed: pellets contain mitochondria/lysosomes
Autoradiography
- Using a radioisotope to track or identify selects protein synthesis (35S-methionine), protein kinases (ATP32) - Label cells → gel → image the gel - Ex. what proteins are selectively synthesized in HeLa cells in response to insulin?
hESCs/stem cell therapy
- hESCs form "embryoid bodies" in vitro - Stem cell therapy requires cell culture
Cell lines
- immortal; divide forever, cancer research, unlimited # of cells, good for purifying proteins, critical for biologics like mABs - sources: 1. Select embryonic cells 2. Cancer cells 3. Telomerase-immortalized cell line: cell strain converted to cell line, manipulate with telomerase
Cell strains
- most physiologically similar to native tissue, comes directly from tissue; often originates from a tissue, divide a finite # of times, con: they die in culture and are limited in amount - Protocol: 1.Tissue separates using EGTA 2.Cell separation 3.Culture: hard to keep this sterile, primary cell culture splits into secondary cell cultures
siRNA/shRNA/antisense RNA
- siRNA = ADV: clinical trials, simple, fast DIS: temporary, non-renewable, not easily transfected into cell line - shRNA = ADV: transfection delivery, permanent, DIS: challenging, difficult design - Target = mRNA → cut into pieces
Electron tomography
-Life in 3D from 2D - Montage of a computer based 3D reconstruction of an EM tomogram with a LM-fluorescence image of the cells (green) on a carbon coated EM grid & Immature HIV Virus - Technique for obtaining detailed 3D structures of subcellular macro-molecular objects - Uses a transmission electron microscope to collect the data.
Intracellular injection techniques (4 different types)
-Neurons and other cells - i.e. Lucifer yellow - Design: fluorescent, vital dye, diffuse through cell, membrane insoluble - Demonstrates cell to cell connectivity through gap junctions
Micro spectrofluorometry/Cytofluor
-Qualitative: microscope (images) -Quantitative: plate reading - Spectrofluorometer = "cytofluor" - Used within cell culture to analyze vital fluorescent dyes
Scanning electron microscopy
-Resolution is 10x less than TEM - Surface structure - Ruled by Abbe's equation
Transmission electron microscopy
-TEM - Internal structure - Beam of electrons is transmitted through a specimen to form an image - The specimen is most often an ultra thin section less than 100 nm thick or a suspension on a grid - High resolution
Fetal bovine serum/defined media
Contains growth factors but serum-free medium is preferred for clinical applications, the serum has a low level of antibodies, which allows it to be used in many different cell culture applications. However, this technique is not efficient for stem cell therapy because there is an increased probability for contamination by viruses and/or harmful bacteria which may compromise the host by producing an adverse immune response
Deconvolution Microscopy
Deconvolution microscopy uses an algorithm to create a 3D view of fluorescently stained cells
Scanning Tunneling microscopy
First that could image DNA molecule
Ex. of use bright field microscopy
Retina, Thyroid gland, Bald scalp, Basal cell carcinoma
Ex. of spinning disk microscopy
cochlea, cell division in embryos (spinning disk), pollen grain (optical sections) and root tip