MBIO 4010 Immunology Lab

MBIO 4010 Teaching Assistant:
Jennifer Tanner and Aniel Moya Torres

All lab reports must have a completed Honesty Declaration attached: Group Lab Assignment or Individual Lab Assignment. Available as pdf.

Lab starts Thursday, January 12, 2012 in rooms 201 and 204 Buller (Possibly 302). Lab manuals are handed out in class the first week of lectures. All essential lab related information is given in the lab manual.

Read lab report presentation section in general instructions at the beginning of your lab manual before doing your lab report. Lab reports must be typed. This includes calculations. 10% of the mark subtracted for portion of report not typed.

Report report formats (Word): Lab 1, Lab 2, Lab 3, Lab 4 and Lab 5 question. First save the file then open in Word to add required information. Correction to lab 2- question 3 now agrees with asterisks in question 2 tables. Refer only to report format not lab manual report.

Report report formats (Excel): Lab 2 and Lab 5. First save the file then open in Excel to add required information.

Lab 4 Hemagglutination Results: All pictures are taken with A1 in the top left corner.group 1, group 2, group 3, group 4, group 5, group 6, group 7, group 8, group 9, group 10, group 11, group 12, group 13, group 14, group 15, group 16, group 17, group 18, group 19, goup 20, group 21, group 22, group 23.

Lab 5 Individual CV% Accuracy and group ELISA data .

Lab References (Not all reference links are used every year, vary depending on report questions.)
Majority of articles available on-line. If link does not work, use Goggle Scholar through UM eLibrairies.

To access some of the journals you need to use UM ejournal, then select Goggle Scholar to search. For example, Annu. Rev. Genet. article.

Biological Safety Cabinet Manual - Labconco Purifier CELL Logic

Lab 1
Anthony R. Newcombea, Chrissie Cresswella, Susannah Daviesa, Keith Watsonb, Guy Harrisb, Kieran O’Donovana & Richard Francisa. 2005. Optimised affinity purification of polyclonal antibodies from hyperimmunised ovine serum using a synthetic Protein A adsorbent, MAbsorbent® A2P. Journal of Chromataography B 814: 209-215
Coffinier Y, Legallais C, Vijayalakshmi MA. 2002. Separation of IgG from human plasma using thiophilic hollow fiber membranes. J. Membrane Science 208: 13-22.
Evtushenko, M, Wang, K, Stokes, HW, Nair, H. 2005. Blood protein purification and simultaneous removal of nonenveloped viruses using tangential-flow preparative electrophoresis. Electrophoresis. 26: 28-34.
Castilho, LR, Anspach, FB, Deckwer, W-D. 2002. Comparison of affinity membranes for the purification of immunologlobulins. J. Membrane Science 207: 253-264. via UM NetDoc Science Direct, UM ejournal, etc.

T.M. Thomas, E.E. Shave, I.M. Bate, S.C. Gee, S. Franklin, D.B. Rylatt. 2002. Preparative electrophoresis: a general method for the purification of polyclonal antibodies. J. Chromatography A 944: 161-168. via UM ejournals

Lab 3
CLIFFORD L. HOWELLt, MARJORIE J. MILLER, AND WILLIAM J. MARTIN. 1979. Comparison of Rates of Virus Isolation from Leukocyte Populations Separated from Blood by Conventional and Ficoll-Paque/Macrodex Methods. J. Clinical Micro. 10: 533-537.
Isolation of granulocytes from human peripheral blood by density gradient centrifugation. MAC Miltenyi Biotec protocol.
Blood cell types (Through the Microscope Blood Cells Wadsworth Centre New York State Department of Health) http://www.wadsworth.org/chemheme/heme/microscope/celllist.h

Lab 4
SHYAM SUNDAR,* SHRINKHLA AGRAWAL, KALPANA PAI, MICHAEL CHANCE, AND MARCEL HOMMEL. DETECTION OF LEISHMANIAL ANTIGEN IN THE URINE OF PATIENTS WITH VISCERAL LEISHMANIASIS BY A LATEX AGGLUTINATION TEST. Am. J. Trop. Med. Hyg., 73(2), 2005, pp. 269–271.
Xu X, Jin M, Yu Z, Li H, Qui D, Tan Y, Chen H 2005. Latex agglutination test for monitoring antibldies to avian influenza virus subtype H5N1. J. Clincial Micro 43: 1953-1955.
Meijer A, Bosman A, van de Kamp EEHM, Wilbrink B, van Beest Holle, MDR & M Koopmans. 2005. Measurement of antibodies to avian influenza virus A(H7N7) in humans by hemagglutination inhibition test. J. Virological Methods 132: 113-120.
Tom van Gool, Hans Vetter, Tony Vervoort, Michael J. Doenhoff, Jose Wetsteyn, and David Overbosch. 2002 Serodiagnosis of Imported Schistosomiasis by a Combination of a Commercial Indirect Hemagglutination Test with Schistosoma mansoni Adult Worm Antigens and an Enzyme-Linked Immunosorbent Assay with S. mansoni Egg Antigens. J. Clin. Microbiol. 40: 3432-3437.
Hull-Jackson, C, Glass, MB, Ari, MD, Bragg, SL, Branch, SL, Whittington, CU, Edwards, CN, Levett, PN. 2006. Evaluation of a commercial latex agglutination assay for serological diagnosis of Leptospirosis. J. Clin. Micro. 44: 1853-1855. [ASM journal, use UM NetDoc on campus or log-on off campus, ejournal as recent article.]
Cheng AC, O’Brien M, Freeman K, Lum G, Currie BJ. 2006. Indirect hemagglutination assay in patients with melioidosis in northern Australia. Am. J. Med. Hyg. 74: 330-334.

Lab 5
Parisa Badiee, Parivash Kordbacheh, Abdolvahab Alborzi, SeyedAli Malekhoseini, Farideh Zeini, Hosein Mirhendi, and Mahmood Mahmoodi. Prospective Screening in Liver Transplant Recipients by Panfungal PCR-ELISA for Early Diagnosis of Invasive Fungal Infections. LIVER TRANSPLANTATION 13:1011-1016, 2007.
Jong-Wan Kim, Jung-Hyun Shim, Joo-Won Park, Won-Cheol Jang, H.K. Chang, Il Han Song, Sun-Young Baek, Seok-Ho Lee, Do-Young Yoon, and Sue-Nie Park.  2005. Development of PCR-ELISA for the Detection of Hepatitis B Virus x Gene Expression and Clinical Application. J. Clin. Lab. Analysis 19: 139-145.
Birmingham, N, Payankaulam, S, Thanesvorakul, S, Stefura, B, Hayglass, K, Gangur, V. 2003. An ELISA-based method for measurement of food-specific IgE antibody in mouse serum: an alternative to the passive cutaneous anaphylaxis assay. J. Immunol. Meth. 275: 89-98. [Available through UM ejournals, just search the J Immunol Methods or Google Scholar.]
Hong, Y, Berrange, ME, Liu, T, Hofacre, CL, Sanchez, S, Wang, L, Maurer, JJ. 2003. Rapid Detection of Campylobacter coli, C. jejuni, and Salmonella enterica on Poultry Caracasses by Using PCR-Enzyme-Linked-Immunosorbent Assay. Appl. Environ. Micro. 69: 3492-3499.
Lewkowich, IP, Campbell, JD, and KT HayGlass. 2001. Comparison of chemiluminescent assays and colorimetric ELISAs for quantification of murine IL-12, human IL-4 and murine IL-4: chemiluminescent substrates provide markedly enhanced sensitivity. J Immunol Methods, 247(1-2): 111-8.
Lewkowich, IP, Rempel, JD, HayGlass, K. 2005. Prevention of allergen-specific, Th2-biased immune reponses in vivo: Role of increased IL-12 and IL-18 responsiveness. J. Immunol. 175: 4956-4962.
P. Scheu, A. Gasch and K. Berghof. 1999. Rapid detection of Listeria monocytogenes by PCR-ELISA. Letters in Applied Microbiology. 29:416-420.
The biological safety cabinet, class II

Immunology Advance Techniques In-lab Assignment
Date: March 8, 2012
Time: 2:30 - 4:00 pm
Group Size: two lab groups work together
Journal Article: Bruno Martin, Keiji Hirota, Daniel. Cua, Brigitta Stockinger, and Marc Veldhoen. Interleukin-17-Producing gd T Cells Selectively Expand in Response to Pathogen Products and Environmental Signals. Immunity 31, 321–330, August 21, 2009.
Aditional Reference Information:
Introduction to flow cytometer by Misha Rahman AbD seroTec
Invitrogen flow cytometer tutorial (flashplayer)

Reponsible for understanding:
-the information presented in the journal article, especially methods and results used by the authors. Emphasis is on understanding methods and FACS result presentation.
-the operation, principle, data analysis and application of the flow cytometer in combination with fluorescence-activated cell sorting (FACS).

Some possible question types:
-analysis of FACS data in journal article
-forward scatter (FSC) and side scatter (SSC)
-"gating"
-given data present schematic plot either histogram or dot plot
-flow cytometer operation (diagram) - flow cytometer used in the journal article compared to basic flow cytometer
-questions specific to figures and tables presented in journal article
-questions relating flow cytometer and research paper
-procedure questions.
Bring:
-journal article
-any other reference material (best on laptop to use as reference)
-wireless internet (Wi-Fi) is not available, must come prepared with all necessary information previously uploaded to your laptop (optional). No internet capable devices permitted, such as I phones, blackberries, etc.
-non-programmable calculator
-scrap paper
-lab manual

Faculty of Science Cheating, Plagerism,etc

Frequently information is available as a pdf file requiring Adobe Acrobat Reader. Adobe abcrobat reader is available at UM Software ExpressUM Software Express.

Lab Skill Sheet- available for students to use for job applications, just check off the highlighted boxes for the lab course(s) you have taken.
pdf or excel (prior to Fall 2007)
pdf or excel (starting Fall 2007)

email le_cameron@umanitoba.ca
Emails: subject must contain course number and subject, eg. 4010 lab 1 report. If no subject given, email is deleted. Emails replies occur only during working hours. Email must include student name.

phone 474-6542
Department of Microbiology
Buller Bldg
Winnipeg, MB
R3T 2N2