First login post
--z3288970 15:13, 8 March 2012 (EST)
First cell bio lab says hi! --z3288970 15:24, 8 March 2012 (EST)
Wk2 --z3288970 15:29, 8 March 2012 (EST)
Wk3 --z3288970 14:11, 15 March 2012 (EST)
Wk4 --z3288970 14:14, 22 March 2012 (EST)
Wk5 ----Z3288970 15:35, 29 March 2012 (EST)
Wk6 -- forgot to sign in, though i was here.
Wk7 ----Z3288970 14:11, 19 April 2012 (EST)
Wk8 --forgot to sign in
Wk9 --Z3288970 14:13, 3 May 2012 (EST)
Wk10 --Z3288970 14:30, 10 May 2012 (EST)
Wk11 --Z3288970 13:54, 17 May 2012 (EST)
Wk12 --Z3288970 14:04, 24 May 2012 (EST)
--Z3288970 14:09, 31 May 2012 (EST)
[membrane AND mboc4[bookPlasma membrane search in an online textbook]]
- Inverted microscope layout
- Solves focal length problems, allows special optical techniques
- Confocal microscopy
- Laser illumination & Fluorescence emission
- Pinhole filter in the same focal plane as the sample = “Confocal”
- 'Scanning' vs 'Spinning Disk' - Methods of scanning of the sample, spinning disk is faster.
- Multi-illumination - usage of laser illumination allows multiple light sources.
- Total Internal Reflection Fluorescence microscopy
- Is very useful to studying the interaction of a cell with a surface
- Live cell imaging
- Must keep cell in ‘normal’ conditions
- Laser Micro-dissection
- Use a laser and a vacuum to capture/remove cells
- Super-resolution microscopy
- Takes a series of ‘masked images’, slightly shifted to each other, compares them and produces a far higher resolution than the diffraction limit would otherwise allow.
1. Identify a reference article that uses the "superresolution" microscopy technique.
Superresolution imaging of chemical synapses in the brain.
2. What did the paper show that normal microscopy could not show.
Using superresolution microscopy techniques researchers were able to study features down to a nanometer scale, in colour and in 3D. This could not be done with EM techniques nor florescence techniques.
- to reinforce tissue structure
- to enhance staining
- to prevent auto-digestion
- to prevent microbial growth
- It is important to be well aware of the hazards associated with the chemicals you are working with
General precautions - working with biological samples demands a degree of caution
Methods of Fixation
- Fresh frozen
- Fast, retains protein function and structure & fat
- Requires specialized equipment, produces thicker sections, tissue may distort while cutting, freezing then thawing degrades tissue.
- Not preservative
- using organic solvents
- fast, preservative, increases cell permeability
- does not preserve 3d structure, dehydrating
- Aldehyde cross-linked
- Prevents the degradation of protein and nucleic acids - may interfere with protein function and epitopes
- popular in Immunochemistry
- Extremely strong fixative
- Detergents - selectively remove components from the sample
- Isotonic solutions - ant solution outside this range will introduce artefacts
- Cell cultures - very easy to fix
- Paraffin embedding - improved sectioning and preservation
- Cryoembedding - not suitable for large sections of tissue
Lab Three Quiz
Leukocyte extravasion can be a pathogenic process when signaling goes wrong, such as in cases of multiple sclerosis where there is a recruitment of leukocytes into the CNS where they should not be normally present  The presence of excess leukocytes in the CNS blood supply is thought to linked to Disseminated Intravascular Coagulation, or Sepsis
Furthermore, some pathogenic organisms can manipulate or counter the usual signaling processes involved in Leukocyte extravasion an example of which includes Bacilus anthracis more commonly known as Anthrax by which the organism reduces the recruitment of leukocytes to an infection
One of the signalling factors, or chemotactic agents for guiding Leukocyte extravasion is Von Willebrand Factor, released when endothlial cells are injured. 
Group project: Added a very brief introduction and extended the section on the pathway involved.
Musashi-1 Rabbit Antibodies
- [Anti-Musashi-1 by Chemicon (Millipore) - Rabbit polyclonal antibody to Musashi-1, useful for mouse, rat and human specimens and compatible with western blotting and immunohistochemistry. 39kDa]
- [Musashi-1 (D46A8) XP® Rabbit mAb, compatible with Immunofluorescence(Frozen) techniques 1:200 dilution recommended for Immunofluorescence (Frozen)techniques]
- "Musashi-1 and Musashi-2 are RNA-binding proteins which play a role in asymmetric cell division of ectodermal precursor cells by regulating the translation of target mRNA. Both family members augment Notch signaling and repress the translation of m-Numb, a protein that positively modulates differentiation of neural stem cells into neurons. Thus, Musashi contributes to the maintenance of neural stem cells . While Musashi-1 is frequently used as a marker for proliferating neural precursor cells, it is also expressed in epithelial stem cells including intestinal and mammary gland stem cells ."
Anti-rabbit IgG with Alexa Fluor® for Immunofluorescence
--Mark Hill 13:31, 17 May 2012 (EST) You have not completed the peer assessment process yet. If you have made comments on each project page they need also to be pasted here today for me to include in your individual assessment.
Lab 12 - Microarray
Identify a current technique used in gene sequencing. Identify a recent cell biology research paper that has used microarray technology. What aspect of the research findings were contributed by the microarray technique.
- High-thoroughput DNA sequencing: By parallelising the DNA squencing process, new generation machines are able to vastly increase their thoroughput.
- Identification of MicroRNAs Inhibiting TGF-β-Induced IL-11 Production in Bone Metastatic Breast Cancer Cells
- Microarrays were used in the aforementioned study to assess the type and levels of miRNA present in the samples produced in the study.