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From CellBiology

Lab attendance

--Nathan Weller 10:42, 10 March 2011 (EST)

--Nathan Weller 09:11, 17 March 2011 (EST)

--Nathan Weller 09:54, 24 March 2011 (EST)

--Nathan Weller 09:23, 31 March 2011 (EST)

--Nathan Weller 09:09, 7 April 2011 (EST)

--Nathan Weller 10:45, 14 April 2011 (EST)

--Nathan Weller 09:22, 21 April 2011 (EST)

--Nathan Weller 09:23, 5 May 2011 (EST)

--Nathan Weller 09:23, 19 May 2011 (EST)

--Nathan Weller 09:14, 26 May 2011 (EST)

--Nathan Weller 09:15, 2 June 2011 (EST)

Lab 1 questions=

1) What are the key cell biology journals? 
  • Journal of cell biology
  • Nature of cell biology
  • Trends in cell biology
  • Cell
  • Public library of science
2) Which journals allow you to reuse their published content
  • PLOS
  • Journal of cell biology
  • BMC cell biology

Work area

Here is some bold text

Here is some italic text

Intro lab

Cell bio

Lab 2 questions

--Mark Hill 08:00, 24 March 2011 (EST) Hi. WHere are your lab 2 answers?

  1. Which chromosomes contribute to the nucleolus?

Chromosomes 13 14 15 21 22 contribute to the nucleolus which is the site for ribosomal RNA transcription.

  1. Identify and add a link to your page of a recent cell biology article using confocal microscopy from the Pubmed database.
  • "Confocal imaging from deep brain tissue" [1] |

Work area

Lab 3 questions

1) "Safety data sheet for chloroform" http://msds.chem.ox.ac.uk/CH/chloroform.html%7C

Has been noted as cancer causing in laboratory animals and is also a carcinogen in humans. Exposure through inhaling or ingestion is toxic and can be fatal. Dermatitis can be caused by exposure to the skin.

2) Pic 2.jpg

Lab 4 Questions

1. Identify a commercial supplier of an antibody that relates to your group project topic.

Abcam is a supplier of antibodies relating to synaptic junctions. They produce Piccolo antibody-A synaptic marker. [2]

2. In mitochondria, where is the gene located that encode Cytochrome C and what keeps this protein trapped within the mitochondria?

The gene that encode Cytochrome C is located in the nucleus of the cell. This protein is a part of the electron transport chain location in the inner membrane of the mitochondria. This protein is soluble in water and is contained by the outer membrane of the mitochondria.

Lab 6 questions

Phenotype.JPG


1 a) What are the changes in phenotypes that you observe between group A and B?

Graph

Group A contains Tm4 and Group B is the control.

-Phenotype A shows no Tm4. -Phenotype B and C are similar in ratio with less Tm4 compared to the control. -Phenotype D and F shows a significant increase in Tm4 compared to the control. -Phenotype E shows a slightly higher amount of Tm4 than the control.

Pictures

Group A there is a generally pink appearance with yellow edges and blue nuclei. Prolonged phenotype and cells are likely to be close together.

Group B they are red with purple nuclei and fanned phenotype. The cells are bound together.

b) How does Tm 4 mediate these changes Tm4 is an actin-binding protein and is a stabilizer in the cytoskeleton of non muscle cells. It has mediated the differences in the two groups as it has caused a prolonged phenotype in the cells in A whereas there is a more fan like appearance in B. This is because the Tm4 has stabilized the actin filaments in the cells in Group A hence the prolonged phenotype.


Lab 9

1. Identify from one of the cell line repositories: a neural cell line and a muscle cell line.


Cell line products from the ATCC:

Neural Cell Line: CRL-2768 ATCC:CRL-2768

Muscle Cell Line: CRL-197 ATCC:CCL-197


2. Identify the species and growth conditions for these cell lines.


Neural cell line: CRL-2765[16]

Species: Rattus norvegicus (rat)

Source: cell type-neuronal Schwann cell; spontaneous immortalization

Morphology: neuronal

Growth Properties: adherent


The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium. To make the complete growth medium, the following components should be added to the base medium: fetal bovine serum to a final concentration of 10%.


Growth condition:

Air 95%; carbon dioxide (CO2) 5%

37.0°C (temperature)

Remove and discard culture medium.

Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.

Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.


Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.

Add appropriate aliquots of the cell suspension to new culture vessels.

Incubate cultures at 37°C.

A subcultivation ratio of 1:6 to 1:10 is recommended

Medium renewal, 2-3 times weekly

Preservation; freeze medium; complete growth medium supplemented with 5% (v/v) DMSO

Storage temperature: liquid nitrogen vapor phase

recommended serum:ATCC 30-2020


Muscle cell line: CCL-198 [17] from ATCC [18]

Species: Mus musculus (mouse)

Morphology: fibroblast

Growth Properties: adherent

Source: Tissue- skeletal muscle, Cell type- fibroblast

Growth condition:

To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.

A subcultivation ratio of 1:2 to 1:5 is recommended

Medium renewal - twice per week

Remove medium, add fresh 0.1% trypsin for about 10 minutes, and centrifuge to remove trypsin. Resuspend cell pellet in fresh medium and dispense into new flasks. Subculture every 7 days. [edit] Peer assessment comments


Peer review

2)

• The page goes very in depth into the topic showing a great understanding of it.

• Comparisons done using the table with other junction and the use of sub headings gave a great flow to the page. This allowed for an easier understanding of the content for someone using it as a learning tool.

• The pictures in the disease section are used very well and tie into the area. The way they are used to space the individual deiseases apart makes the section easy to digest and take in.

• The only thing that looks out of place are the pictures in the structure section, maybe place them to the side of the text so they aren’t disconnected from it.

• Some stats and treatments for the disease section can be added.

• Glossary could use more content.


3)

• The intro and history are both good as they provide a broad beginning to set up the page.

• Pictures are well positioned.

• The content in the structure section is fine but the texts location could be changed so it all fits together better.

• Molecular components provides in depth knowledge but was difficult to get through and learn, a table could make this easier to digest.

• Functions was well structures with headings and the diseases table was spot on.

• Future research is needed perhaps on research into the cures or treatments for some diseases.


4)

• Even though the intro is short, it still addresses the topic well.

• The page overall was in depth in respect to knowledge of the topic and a lot of research has obviously been done. However trying to learn from some of the sections with so much bulk can be difficult.

• The pictures tied in well to the areas but there size can be increased.

• Regulations can be decreases in size using a table.

• Any research being carried out in something on this topic other than diseases.


5)

• A very well structures page, I found it easy to understand throughout and the balance between too much and not enough content was found.

• A brief yet informative intro and history sets up the page well, and the picture is clear and informative.

• The structure shows in depth research.

• Diseases, types and current research is good as it is clear and easy to follow.

• A diagram for function may be needed to explain it further.


6)

• Intro and history along with an expertly drawn diagram sets the page up very well.

• After reading I don’t think anything to do with neuromuscular junctions has been left out, obviously a lot of work and research has been done here.

• The page was very easy to read and understand due to the use of point form and well placed pictures.

• Tables in the dysfunctions area is especially good.

• Only current associated research was what I found to be bulky and dense otherwise a perfect page to learn about the topic.