User:Z3422484: Difference between revisions
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==Lab 1 Assessment== | |||
<pubmed>25100708</pubmed> | |||
The article above investigates in detail, the acrosome reaction occurring within spermatozoa and whether other roles are involved with [Ca2+]i. This was facilitated by using FM4-64, Fluo-4 AM (fluorescent dye) and Sytox green (represent whether cell death has occurred) as well as progesterone, Ham's F-10 media, FITC-PSA and ionomycin. The three major steps undertaken in the experiment (method) involve the preparation of the human sperm samples, viewing of spermatozoa samples and the use of the scanning electron microscope. The preparation of the sperm samples involved culturing sperm in a Ham’s F-10 medium in an incubator at room temperature and then placed into medium allowing capacitation to occur. Spermatozoa were then suspended in various reagents including the FM4-64 DYE, incubated then viewed under an inverted microscope. The spermatozoa (given progesterone) were then viewed under a scanning electron microscope. | |||
The study found that applying ionomycin with the FM4-64 dye led to a more accurate image forming on the acrosomal region of spermatozoa due to an increase in fluorescence. With this in mind, distribution changes occurred when AR inducers (ionomycin and progesterone) were applied. The acrosomal reaction is heavily dependent on extracellular Ca2+. The staining revealed the formation of tube-like structures on the spermatozoa head. These structures were unique in that developed while capacitation was underway. Also, it was found that progesterone did not affect spontaneous Ca2+ oscillation and therefore no correlation was found. |
Revision as of 15:49, 12 August 2014
Welcome to the 2014 Embryology Course!
- Links: Timetable | How to work online | One page Wiki Reference Card | Moodle
- Each week the individual assessment questions will be displayed in the practical class pages and also added here.
- Copy the assessment items to your own page and provide your answer.
- Note - Some guest assessments may require completion of a worksheet that will be handed in in class with your student name and ID.
Individual Lab Assessment |
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|
Lab 12 - Stem Cell Presentation Assessment | More Info | |
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Group | Comment | Mark (10) |
1/8 |
|
7 |
2 |
|
7.5 |
3 |
|
7.5 |
4 |
|
8.5 |
5 |
|
8.5 |
6 |
|
8.5 |
7 |
|
7.5 |
LAB ATTENDANCE
--Z3422484 (talk) 12:51, 6 August 2014 (EST)
http://www.ncbi.nlm.nih.gov/pubmed
<pubmed>25084016</pubmed>
Lab 1 Assessment
<pubmed>25100708</pubmed>
The article above investigates in detail, the acrosome reaction occurring within spermatozoa and whether other roles are involved with [Ca2+]i. This was facilitated by using FM4-64, Fluo-4 AM (fluorescent dye) and Sytox green (represent whether cell death has occurred) as well as progesterone, Ham's F-10 media, FITC-PSA and ionomycin. The three major steps undertaken in the experiment (method) involve the preparation of the human sperm samples, viewing of spermatozoa samples and the use of the scanning electron microscope. The preparation of the sperm samples involved culturing sperm in a Ham’s F-10 medium in an incubator at room temperature and then placed into medium allowing capacitation to occur. Spermatozoa were then suspended in various reagents including the FM4-64 DYE, incubated then viewed under an inverted microscope. The spermatozoa (given progesterone) were then viewed under a scanning electron microscope.
The study found that applying ionomycin with the FM4-64 dye led to a more accurate image forming on the acrosomal region of spermatozoa due to an increase in fluorescence. With this in mind, distribution changes occurred when AR inducers (ionomycin and progesterone) were applied. The acrosomal reaction is heavily dependent on extracellular Ca2+. The staining revealed the formation of tube-like structures on the spermatozoa head. These structures were unique in that developed while capacitation was underway. Also, it was found that progesterone did not affect spontaneous Ca2+ oscillation and therefore no correlation was found.