CH5 EMBRYO LEC

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1. Dissecting - provides a three dimensional image which 11. Excitation how the intensity of fluorescence varies with
microscope allows accurate perception of depth by the spectrum the excitation wavelength
observer
12. Emission how the intensity of emitted fluorescence is
spectrum distributed across the wavelength spectrum
- assists in the performance of manipulations
such as microsurgery or microinjection 13. Dark-field - depends on illumination from a very
microscopy oblique angle so that only points within the
-does not invert the image specimen that scatter light extensively are
visible
2. Incident used when specimens are opaque (ex. chick
lighting embryos)
- appear as bright points while the remainder
3. Fiberoptic used in living organisms of the specimen is dark
light guide
14. Polarization - places the specimen between crossed
4. Transmitted used when specimens are transparent (ex. microscopy polaroid filters
light base zebrafish, sea urchin, mouse)
- if contains components that can alter the
5. Compound - used for examination of sections or for
plane of polarization of the light beam, it
microscope whole organisms that are small enough to be
appear as bright patches and show
transparent
birefringence
- invert the image 15. Conventional limited to use on sections or very thin whole
fluorescence mounts because fluorescence from cells
6. Ordinary used to examine stained sections or
microscopy above and below the plane of focus would
transmitted wholemounts
otherwise swamp the image
light
16. Confocal - fluorescence from thicker wholemounts can
7. Phase - technique that converts small difference of
scanning readily be visualised by this
contrast refractive index within the specimen into large
microscope
microscopy differences of light intensity
- uses a laser for illumination
- favored method for examining live cells
- illuminates just one point at a time
cultured in vitro or isolated gametes
17. Multiphoton - provide even better optical sectioning of
- used for small transparent embryos such as microscope fluorescent specimens than the confocal
the mammalian preimplantation stages microscope
8. Differential - converts small differences of refractive index
- light source is of less than the enrgy
interference into an apparent difference in height when
required to excite the fluorochrome
contrast perceived by the observer
(Nomarski 18. Charge image collection is done by this device
optics) - provides a sharp resolution of a particular Coupled
optical section within the specimen Devices
(CCD)
- provides very clear visualization of single
19. Most - function only in black and white
cells within small transparent specimens
sensitive
CCD - can collect long exposures from very faint
- technique that enabled the elucidation of the
specimens
entire cell lineage of the worm C. elegans
9. Fluorescense used to visualise the location of a flourescent - able to count single photons
microscopy substance (fluorochrome) within a specimen
20. Less - collect images fast enough to display on a
10. Principle of The principle of fluorescence is that the sensitive video screen
fluorescence fluorochrome absorbs light of a particular CCDs
energy and emits light of a lower energy. - enable the recording of movies in real time
Lower energy = lower wavelength or a color
21. Time-lapse useful for the study of morphogenetic
shifted toward the red end of spectrum.
imaging movements of cells, organ cultures, or
embryos
22. Cryostat a microtome operating in a cooled 33. Microarray - enable examination of large
chamber numbers of gene products
simultaneously
23. Electron provides more magnification than is
microscopy required to identify patterns of cell types
- identify the genes likely to be
in tissues or embryos
involved in a particular system or
24. Scanning provide vivid three-dimensional views of process
electron wholemounts and is often used for
microscope visualizing cell arrangements in the - often used to make comparisons, for
course of morphogenic movements example between two embryonic
25. Biochemical give a reasonably accurate quantitative stages, or between cells treated and
methods measure but no anatomical information untreated with an inducing factor

26. In situ methods give accurate anatomical information but 34. cDNA arrays - consist of a regular arrangement of
limited quantification closely spaced dots, each of a
different cDNA arranged in a
27. Laser capture molecular analysis on specific cells or
rectangular array on a glass slide
regions of cells taken from microscope
sections
- long synthetic oligonucleotides are
28. Reverse - standard method for detecting and used
transcription measuring RNA
35. Affymetrix system - a proprietary product
polymerase
chain reaction - examines the accumulated concentration
- larger number of short
(RT-PCR) of product at the end of the reaction
oligonucleotides are synthesized
directly on the sides
- method is best only semiquantitative
- several oligonucleotides will
- very prone to artefacts and careful
represent sequences from one gene
controls need to be done to ensure
specificity 36. Fluorimeter measure the fluorescence from the
dye bound to each spot
29. Real-time PCR - achieve more accurate quantitative
measurement of specific mRNA levels 37. RNA-Seq ability to measure the composition of
the entire transcriptome in a cDNA
- rate of information of the amplified sample
product os monitored in real time 38. Proteomics identify individual unknown proteins
30. SYBR Green fluoresces brightly when bound to from a complex mixture and the set of
double-stranded DNA but not when techniques employed to do this is
bound to single-stranded DNA referred to as _________

31. Ribonuclease most sensitive method for the detection of 39. Mass spectrometer ionize and volatilize the substance
protection specific mRNA before the adoption of RT- then determine its time of flight to the
analysis PCR detector after acceleration in an
electric field
32. Northern oldest and least sensitive technique
blotting 40. Western blot method that shows the total proteins
- can show the number and size of mRNA of the sample that are extracted and
where there are several splice variants separated on acrylamide gel
formed from the same gene 41. Immunoprecipitation - method of isolating the protein
recognized by a specific antibody

- used to find whether a particular

protein is being newly synthesized in
an embryo or tissue sample

- used in nonradioactive mode to

establish the presence of a particular
protein in a sample
42. Chromatin - combined with microarray analysis 50. Green - protein showing intense green fluorescent
immunoprecipitation so that thousands of loci can be fluorescent emission
(ChIP) examined simultaneouly (ChIP-Chip) protein (GFP)
- often remains active when fused to other
- it has also been combined with deep proteins
sequencing to enable the whole
genome to be examined (ChIP-Seq) - easy to visualize in living specimens and
so allow real-time examination of the
43. In situ hybridization reveals the regions of a specimen
labeled cells as well as detection in fixed
where a specific mRNA is present
specimens
44. Epitopes particular parts of an antigen
51. Luciferase enzyme that catalyzes breakdown of a
molecule that are recognised buy the
substrate (luciferin) in the presence of ATP
antibody
52. Microinjection used to introduce inhibitors such as specific
45. TRUE Enzyme-linked methods are usually
methods antibodies or antisense olignucleotide ito
more sensitive than fluorescence
cells
because the enzyme-substrate
reaction provides an additional 53. Fate mapping show the normal destiny of embryo regions
amplification step. in the course of development
46. Reporter genes - encode some easily detectable 54. Vital dyes -oldest type of extracellular label
product to monitor some particular
55. Fluorescent most commonly used intrcellular labels
aspect of events in the organism
dextrans

- used to analyse the regulatory 56. Retroviruses RNA viruses that carry a reverse
domains of genes transcriptase enzyme

47. B-galactosidase - large, tetrameric enzyme capable of

hydrolyzing a whole range of B-
galactosidases, which are substances
consisting of a chemical group joined
by a B linkage to the 1-carbon of
galactose

- very useful reporter both because of

its sensitivity and because most
animal tissues do not contain cross-
reacting enzymes, which means that
background staining is usually low
48. Perdurance - B-galactosidase enzyme is present
but the mRNA encoding it is absent

- particularly significant in embryos

that do not show much growth
49. B-geo fusion of the B-galactosidase enzyme
with the product of the neomycin
resistance gene and possesses both
of the biological activities in the one
molecule