Lab Methods + Microscopy

growing living cells and isolating from tissues

immortalized hybrid cells created by fusing antibody-producing B cells with myeloma cells (cancerous cells)

used to analyze and characterize cells or other particles in a fluid sample. It allows for the simultaneous measurement of multiple parameters, such as size, shape, fluorescence, and surface markers

Able to separate proteins based on molecular weight (kDA)

used as purification strategies for native proteins

detecting a specific protein in a mixture

identification of macromolecules (usually protein)

rapidly copies specific segments of DNA, creating millions to billions of copies from a small sample

identify cells within a fixed sample that express a certain gene of interest

Quantitative RT-PCR allows accurate quantitative determination of the amount of mRNA in samples

global mRNA analysis

Specifically degrade a target mRNA, then look for effects

Homologous recombination to delete genes

inactivating a specific gene to study its function. Scientists can observe the effects of the missing gene by introducing a mutation, which provides insight into the gene’s role in the organism’s biology.

brightfield, phase contact, DIC, Darkfield

antibodies and fluorescent dyes, used to visualize and localize specific proteins or antigens within cells or tissues

high-resolution imaging technique that uses lasers and a pinhole to block out-of-focus light

class of proteins that emit visible light when exposed to ultraviolet or blue light

advanced optical imaging technique that uses an evanescent wave generated by total internal reflection to selectively excite fluorescent molecules in a very thin layer (around 50–100 nm) at the interface between a glass surface and a liquid

a live-cell imaging technique used to measure the mobility and dynamics of fluorescent molecules within a cell or membrane

biophysical technique that measures distances at the nanoscale (1-10 nm) by observing the non-radiative transfer of energy from a donor fluorophore to an acceptor fluorophore

TEM (Transmission Electron Microscopy) images the internal, cross-sectional structure of ultra-thin samples using transmitted electrons, providing high-resolution, 2D images of atomic structures. SEM (Scanning Electron Microscopy) scans the surface of thicker samples with electrons and detects reflected or scattered electrons to produce a 3D-like surface image with lower resolution than TEM

imaging technique that uses a beam of high-energy electrons to create highly magnified images of the internal structures of samples, achieving sub-nanometer or even atomic-level resolution

high-resolution, three-dimensional surface view by scanning a focused beam of electrons over a sample