Bootcamp Chapter 8 Flashcards
T/F? Samples that are viewed under electron microscopy are not living.
TRUE - samples must be fixed, stained, and killed
What type of microscope allows for the visualization of live cells?
a) bright field microscopes
b) stereo microscope
c) phase contrast microscopes
d) compound microscopes
C
Fluorescence microscopes rely on the _____ of _____ from a particle that has absorbed light for visualization of a specimen
emission of photons
Dichroic Filter
a filter used in fluorescence microscopy that reflect certain wavelengths and allows some to pass through
Distortions in an image in fluorescence microscopy are referred to as ______
artifacts
Which microscope is typically used to observe chromosomes during mitosis?
Confocal laser scanning microscopy
What type of microscopy is used to view UNSTAINED samples of LIVE cells?
dark field microscopy
Scanning electron microscopy contains _____ samples, while cyro-SEM contains _____ samples
dehydrated, frozen
Both types of SEM kill the sample!
Which type of microscope allows for a high resolution 2D image of a samples internal structure?
Transmission electron microscopy (TEM)
You are viewing 3D internal structures of the mitochondria. What process is used to allow this? How does it work?
Electron tomography (not a type of microscope) - done by sandwiching TEM images together
Refer and review tables pages 117-118 in DAT bootcamp
review
A grad student needs to manually count the number of cells in her sample. What technique/material might she use?
Hemocytometers - a gridded slide
Plant cells are being centrifuged for study. What might you expect to pellet (precipitate) first?
The densest and most compact particles
Organize the following constituents from the most dense to least dense
ribosomes, ER fragments, nuclei, mitochondria, chloroplasts
From most dense to least dense - nuclei > mitochondria/chloroplast> ER fragments > ribosomes
The slight difference that are in the human genome occur every 1000 nucleotides. These are referred to as _____ ______ _____.
single nucleotide polymorphisms
Restriction enzymes
enzymes that will cut DNA at palindromic (inverted mirror) sequences to product sticky or blunt ends when making recombinant DNA
Restriction Fragment Length Polymorphisms
Unique lengths of DNA that result from restriction enzymes
What can be used to allow for the comparison between individuals through the polymorphisms in DNA length?
Restriction fragment length polymorphisms - DNA fingerprinting uses this to ID people!
What heat resistant enzyme is used in polymerase. chain reaction?
Taq polymerase
When is the temperature lowered during PCR?
during primer annealing; denaturation starts to split DNA strands but process must cool down to allow RNA primers to anneal
T/F? Bacterial cloning will clone prokaryotic cells in eukaryotic gene products.
FALSE - it will clone eukaryotic gene products in prokaryotic cells!
Vector
a piece of DNA that can be taken up by cells, allowing DNA to be replicated, transcribed, and translated
What process allows for transformation (taking up DNA from the environment) to occur?
electroporation - electricity is applied to the cells, creating holes in the plasma membrane
Gel Electrophoresis
an electric field is applied to agarose gel to create a negative cathode at the top of the gel, and a positive anode at the bottom
In gel electrophoresis, what is being prevented from migrating towards the positively charged anode?
*note - recall that the DNA being inserted in negatively charged!
pores created from electrophoresis will obstruct the movement of larger fragments - smaller fragments will proceed to the bottom
Gel electrophoresis will separate fragments based on ____ and ______
charge and size
Southern blotting is used to ID ______
Northern blotting is used to ID _____ using ____ probes
Which technique utilizes sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) to quantify amount of target protein in a sample?
DNA
RNA, RNA
Western blotting
SNOW DROP
Enzyme Linked Immunosorbent Assay (ELISA)
determines if a specific antigen exists in a person
Pulse Chase Experiments
demonstrate how a protein moves through a cell; gives into about gene expression
What are DNA microarrays typically used for?
determine which genes are expressed and which are not expressed in a type of cell
example - cancer research
In order to a clone an animal via reproductive clone, a somatic cell would need to be reverted from its _____ to its _____ state
multipotent, totipotent
Differentiate between totipotent, pluripotent, and multipoint.
Totipotent - can give rise to an entire organism because it can product extra embryonic membranes
Pluripotent - can give rise to any cell type because it can differentiate into any of the three germ layers
Multipotent - cannot develop into an entire organism and can only give rise to some of the germ layers, but not all
Fluorescence Return After Photobleaching (FRAP)
a type of live cell visualization that allows researches to see how and where biomolecules are moving in a live cell
gives quantitative insight to cell dynamics
Fluorescence Lifetime Imaging Microscopy (FLIM)
gives quantitative measure of the concentration of various ions, molecules, and gases in a cell - achieved by irradiating cell samples with light and measuring their fluorescent lifetime
Knockout Mice
a genetically modified mouse in which researchers have inactivated, or “knocked out”, an existing gene by replacing it or disrupting it with an artificial piece of DNA
Which types of microscopes can view LIVING samples?
dark field, phase contrast, confocal laser scanning, stereomicroscope (light), compound microscope (light), and fluorescence
