1. Microbiology 156 Instructor: Lucia Testa

2. Week 1 WELCOME TO MICROBIOLOGY LAB 156 Syllabus Homework Assignments Study Guide Safety Contract Exp. 4: Microscopy Lab Protocol- Journal Aseptic Routine Exp. 1: Culture Transfer (without organisms) Lab Techniques

3. Safety Routine  This is a microbiology lab—there are lots of germs in here!  Leave your personal belongings in the cubbies provided  No eating or drinking (yuck!)  No cell phones/text messaging  Lab coats in lab only  Long pants and closed-toed shoes; tie your hair back  Wash hands and wear gloves  Disinfect lab table before and after experiment  Throw out waste in proper receptacles.

4. Safety Routine  Coming In 1.Place items in cubby (inc. food/drink) 2.Personal check (coat & hair) 3.Wash lab bench. 4.Wash hands/put gloves on.  Leaving 1.Equipment/ culture away/dispose 2.Wash bench. 3.Dispose of gloves in red biohazard bin. 4.Wash hands.

5. Creating a Lab Journal

6. Laboratory Notebook Protocol primary form of documentation in the research laboratory legal document can be used to establish a patent claim, to assign credit for a discovery, to document data for publication, and document historical data critical that the laboratory notebook be used properly and consistently since the laboratory notebook has legal significance (grading significance for students), the following formatting, rules, guidelines and specifications apply refer to tutorial at biolab156.wikispaces.com website

7. Entry Protocol for Microbiology Laboratory Experiments (Found in Suppliments on pg 2C) Header: Exp. #, Title, Date. Every page shall have the Header with : Exp. #. Title, date Example: Header: Exp. 2 Handling & Examining Cultures June 5, 2002 Purpose: The purpose and/or objective of the experimental activity, hypothesis, and premise. Example: Purpose: To make aseptic transfers of pure cultures and to examine them for gross features. Materials: List cultures, media, & equipment to used for activities. Example: Material: E. coli broth culture, B. cereus slant culture, inoculation loop, (2)TSA broth, (2)TSA plates. Procedures/Activity: briefly describe procedures, reference sections and page numbers of the lab manual for specifics and details. Example: Procedure: *(Cappuccino & Sherman) Microbiology Lab Manual pages 13-14, Obtained cultures from instructor, aseptically transferred cultures per instructor’s inoculation scheme and Exp. 2, to TSA broth, TSA plate, incubated at 37 o C for 24 hrs, refrigerated until next period. Reviewed and recorded results. Result: data, drawing, observations, tables of results of tests, details of results. All diagrams of microscopic field should be 3 inch (5 cm) in diameter and labeled with color and magnification. Make drawing of cultures and/or slides. Make a table of results of tests. Example: Results: record observation of cultures: colors, consistency, odors, appearance. Conclusion: Restate objective, then make statements of fact about the data to support objective, hypothesis, and premise. Summary of results.

8. Journal Notebook Protocol 1 st : Enter you name, class, and information to find you if found on the out side cover for identification. 2 nd : Number every page of book, top right side only. 3 rd : Label the top of the first six pages of the book as : Table of Content. 4 th. Write the complete reference of the Laboratory Manual on the last page of your journal so that you can reference it. All entries are in chronological order by date. All entries are in black pen All entries are made direct to book ( no scrap paper or other notebooks). Entries are made on right hand pages only, left hand pages are for class notes, calculations, instructions. State sources for all literature values or outside sources of information. Every right page is date when the entry is made. Every right page has the Experiment #, Experiment name, and date as a header. Every right page at the end, lower right designates continuance or end. Every right page is initialed when completed. The Table of Contents shall reflect all entry pages by Exp. # and Name with page numbers and dates of the work. Never rip out any pages. Cross out, do not erase or white out mistakes. Use a single strike through line and initial the cross out. Do not leave blank page or blank areas on a page, draw a diagonal line through the unused portion or pages. Tables may be drawn in or taped in from a lab manual or spread sheet. Note: proper credit must be given to the author. Include date of entry. Hand outs with procedures, charts, reference data, etc. may be taped in with the referenced author, book, etc. and date of entry. Spreadsheets may used and be taped in from student developed spread sheets. Note: proper credit must be given to the author. Include date of entry.

9. Experiment 4 Microscopic Examination of Stained Cell Preparations How big...? How big...?

10. Exp 4: Microscopy Review of microscope parts Principles of microscopy Care and operation of microscope Procedures for observation: letter “e” print Procedures to observe cross thread slide with oil immersion objective lens Cleaning and storage Materials: Prepared slide - “e” print Prepared slide of cross threads

11. Refractive Index

12. Getting Started: Exp 4: Microscopic Examination of Stained Cell Preparations Obtain a prepared slide of an “e” and a slide of cross threads. Follow directions described in lab manual on pg 31-36. Scanning (4X) lens first, place slide on. Center the slide and use coarse-adjustment knob to focus. Center the slide and use coarse-adjustment knob to focus. Switch to higher magnifications (slide should still be centered) using fine-adjustment knob to focus. Switch to higher magnifications (slide should still be centered) using fine-adjustment knob to focus. Observe @ 1000 x total magnification with oil immersion technique.

13. Exp 4: Microscopic Examination of Stained Cell Preparations date THIS WEEK AT HOME Purpose: In lab manual (pg 31) Materials: Prepared slides of Gram stained bacilli and cocci organisms, microscope. Procedures: (* Cappucino & Sherman, 9 th ed. Pages 36) bullet procedures, ref page # in lab manual FOR NEXT WEEK IN CLASS Data: drawings (3 inch [5 cm] field of view, details) Conclusion: Pg # Initial

14. Exp 1: Culture Transfer Techniques

15. Getting Started Exp 1: Culture Transfer Techniques (Aseptic/Sterile Technique) Material: Culture: 1 practice culture- mystery microbe [MM] (per table) Media: 1 TS broth, 1 TSA slant, 1 TSA deep (per person) Procedure: Lab One * (Cappuccino & Sherman, 9th ed. Pages 9) Properly label each medium. Aseptically transfer (inoculate) to each medium as described on pg 7-9.

16. For Next Lab period: - Follow WEEK 1 Homework Assignments on page 6 - Read Case Study/ Assignments page 9