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Human Bioscience 2 BMS192 Laboratory Manual Faculty of Science

Written and compiled by Rhett McClean Archana Buttsworth Brian Spurrell Adam Hamlin Gordon Paterson Patricia Logan Kelly Linden Jillian Dunphy Lesley Forward Ingrid Stuart

Produced by Division of Student Learning, Charles Sturt University, Albury - Bathurst - Wagga Wagga, New South Wales, Australia.

Printed at Charles Sturt University © Charles Sturt University Previously published material in this book is copied on behalf of Charles Sturt University pursuant to Part VB of the Commonwealth Copyright Act 1968.

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Contents Page Instructions for practical sessions

1

Human experimentation

1

Ethics in human experimentation

2

Laboratory safety rules

2

Laboratory dress General information Waste disposal

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Hints on anatomical drawing

5

Faculty of Science Laboratories and specialist teaching spaces student induction form

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Personal Protective Equipment (PPE)

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Home exercise 1: Endocrine system

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Introduction The hypothalamic-pituitary axis Thyroid gland The adrenal gland The gonads Practical 1: Blood Introduction Activity 1: Blood groups Activity 2: Haematocrit Activity 3: Haemoglobin content Activity 4: White blood cell identification Practical 2: Cardiovascular anatomy Activity 1: Systemic and pulmonary blood flow Activity 2: Anatomy of the heart Activity 3: Heart dissection Activity 4: Histology Activity 5: Blood vessels Activity 6: Foetal circulation Practical 3: Cardiovascular physiology Introduction Activity 1: Heart sounds Activity 2: Heart rate Activity 3: Measurement of blood pressure Activity 4: Effect of exercise on blood pressure Activity 5: Control of blood pressure

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Activity 6: Conduction system of the heart. Activity 7: Electrocardiography Practical 4: Respiratory system Introduction Activity 1: The respiratory passages Activity 2: Histology Activity 3: Lung dissection Activity 4: Pressure changes during pulmonary ventilation Activity 5: Pulmonary function testing Activity 6: Dynamic lung function tests Activity 7: Control of ventilation: the role of carbon dioxide Home exercise 2: Digestive system Introduction Practical 5: Gastrointestinal physiology Introduction Activity 1: Histology of the digestive tract Activity 2: Digestion of starch Activity 3: Bile emulsification Practical 6: Urinary system Activity 1: Renal anatomy Activity 3: Histology of the urinary system Activity 4: Renal physiology Practical 7: Reproductive system Introduction Activity 1: Male reproductive system Activity 2: Female reproductive system Practical 8: Inheritance Introduction Activity 1: Basic genetic language Activity 2: Monohybrid crosses and the punnet square Activity 3: Incomplete dominance Activity 4: Codominance Activity 5: Sex linked inheritance Activity 6: Additional inheritance problems

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BMS192 Practical Manual

Instructions for practical sessions 1.

Attendance at practical sessions is compulsory. If you are unable to attend a practical class, inform the lecturer as soon as possible, and provide documentary evidence wherever possible (e.g. medical certificate).

2.

Regulations concerning laboratory safety are included in these notes. You should read these regulations carefully and abide by them at all times. Staff will refuse entry to any student wearing inappropriate footwear. You need to wear sturdy shoes which cover the foot and heel.

3.

You should bring your practical notes and text book to all practical sessions.

4.

Leave your place in the laboratory tidy. Wash, dry and put away equipment as appropriate. Replace your seat under the bench when not in use.

5.

Read through the notes before each practical session. Complete as much of the diagram labelling as possible before the practical session.

6.

During the practical session you should complete all the exercises listed and check with a demonstrator before leaving the laboratory.

7.

Organise your practical session so you do not get behind. For example, if there is only thirty minutes to go and you have three pages to complete only spend ten minutes on each page. Practical sessions will finish promptly at the scheduled time.

Human experimentation

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In this subject you will be performing experiments involving human subjects. There are two important issues regarding the use of human subjects that you should be aware of: 

The ethical issues relating to the use of humans as experimental subjects. The issues are outlined in the following section entitled Ethics in human experimentation;



The safety issues associated with working with materials of human origin, such as blood, urine or saliva. The special precautions involved with such biohazardous substances as outline by laboratory staff.

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In some laboratory practical classes, exercises using blood have been devised where the blood samples used are obtained from volunteer staff, students and patients by venepuncture or a finger prick. Other body fluids or tissue (e.g. urine, saliva and cheek cells) may also be utilised.



Some research projects also require the collection and handling of blood and other materials of human origin.

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All samples used by students will be either from screened donors (i.e. showing negative serology/virology for syphilis, Hepatitis B and C, and HIV) or provided by students who will be testing their own body samples themselves. This does not guarantee any samples are disease free.

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

Teaching and research activities that cannot meet their objectives through the use of the types of sample outlined above must obtain approval for an exemption to this policy. The application for an exemption should be made in writing, be addressed to the University Biosafety Committee and should include a detailed risk management protocol.

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No body samples from unscreened donors (with the exception of the samples provided by the students for their self-testing) shall be used unless approval for that specific activity has been provided, in writing, by the Biosafety Committee.



All Human Biological specimens should be regarded as infectious at all times.

Ethics in human experimentation At Charles Sturt University, experiments involving human subjects are conducted according to the guidelines set out by the National Health and Medical Research Council entitled NHMRC Statement on Human Experimentation - copies of these guidelines are available for perusal on request. These guidelines were designed to apply to the conduct of research; however, most of the clauses are relevant to the conduct of undergraduate experiments involving human subjects. Perhaps the most important clause is clause 8, which states: ‘Before research is undertaken, the free consent of the subject should be obtained. To this end, the investigator is responsible for providing the subject (at his or her level of comprehension) with sufficient information about the purpose, methods, demands, risks, inconveniences and discomforts of the study. Consent should be obtained in writing, unless there are good reasons to the contrary.’

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In the laboratory experiments you will perform, clause 8 will be applied as follows: 

your participation as a subject in such experiments is entirely voluntary;

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students who know that they have infectious/contagious diseases should not volunteer;



information about the purpose, risk etc. of laboratory exercises is provided in the laboratory notes;



by agreeing to participate as a subject for such experiments, you are giving your informed consent - no written consent will be obtained.

Laboratory safety rules The following instructions are for your protection. Observe them at all times. Each session students will be inducted into the laboratory by laboratory staff and then sign a student induction form.

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Laboratory dress 

Students must wear closed-in, non-slip footwear. Anyone wearing thongs or open-toed or open backed shoes will not be permitted in the laboratory.



Properly fastened laboratory coats are mandatory but they must not be worn outside the laboratory. Laboratory coats can only be taken from the laboratory if suitably contained in a safe manner.

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Disposable gloves are available for use and are recommended when handling biohazardous substances or if hands contain open wounds.

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Always use safety glasses when necessary or instructed to do so.

General information 

You must obtain your supervisor’s permission and that of the Laboratory technical staff to use the laboratory out of timetabled periods.

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Unauthorised experiments in the laboratory are strictly forbidden.

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When in the laboratory, never adopt a casual attitude - always be conscious of potential hazards. No horseplay or running is permitted in the laboratory.

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Always take note of instructions given by your lecturer, demonstrators or the laboratory staff.

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Regard all substances as hazardous unless there is definite information to the contrary.

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No mouth pipetting is permitted at any time.

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No human venepuncture may be performed by an undergraduate student.

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Report all accidents - no matter how slight - to the lecturer or laboratory staff.

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Familiarise yourself with the locations of all fire exits and extinguishers, and with the safety facilities available in the laboratory.

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There are first-aid kits available in each laboratory and emergency eye-wash facilities in each laboratory.

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Emergency gas cut-off switches are located near the main entrances in each laboratory.

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Do not attempt to adjust instrumentation or microscope light sources if you are not familiar with them. Always ask the laboratory staff for assistance.

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If you notice any faults with any piece of equipment, or any hazardous situation in the laboratory, please report it immediately to the lecturer or laboratory staff.



On completion of your practical session you are to ensure that your work areas are left clean and tidy. If biohazardous materials have been used the

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BMS192 Practical Manual

benches must be wiped down with disinfectant (sodium hypochlorite wash bottles are located in each bay). 

Scalpels should be returned to the distribution point and not placed with other dissecting instruments. Damaged scalpels or scalpel removal is to be done only by Laboratory technical staff.

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All dirty glassware (including test tubes) is to be rinsed and placed in the tubs provided. See special instructions for biohazardous material.

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Always wash your hands before leaving the laboratory.

Waste disposal General waste Paper, plastic and non-contaminated material, etc, can be placed in the plastic lined garbage bins located in each bay. Biohazardous waste Contaminated slides, pipettes, pasteur pipettes etc, will be placed in disinfectant containers located in each work area. Serology tubes, agar plates, dirty gloves and contaminated paper, etc, are to be placed in autoclave bags ‘biohazard waste’ bins located in each laboratory. Sharps

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Needles, scalpel blades, etc, should be placed in the special ‘sharps’ containers located in each laboratory. Glass

Broken glassware, uncontaminated pasteur pipettes and serology tubes, etc, are to be disposed of in the ‘glass only’ bins located in each laboratory. A dustpan and broom is available from the prep room for cleaning up broken glass. Under no circumstances should broken glass be picked up with your hands. Hazardous chemicals Follow instructions given. Waste bottles are placed in the fume hoods for organic, heavy metal and other wastes that are not to be disposed of down the sink. If you have ANY doubts, always ASK your instructor or the laboratory staff.

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Hints on anatomical drawing You do not have to be artistic to make good anatomical drawings. However, you do need to be able to draw accurately the two dimensional outline of a three dimensional object. This also means that you need to be able to recognise the important features of a given microscope slide, organ or whatever. The following hints should help you make accurate and meaningful records or your laboratory observations. 1.

Use a medium grade pencil (HB or H only).

2.

Draw only what you see (not what you think should be there).

3.

Make large diagrams (leave only enough room around the margins for labelling).

4.

Give every diagram a specific title – including the section and/or direction of view.

5.

Give magnification (eyepiece magnification x objective magnification) where appropriate.

6.

Give source e.g. Marieb (1998) text, model, microscope slide etc.

7.

Label all relevant parts of diagram.

8.

Draw a small area accurately rather than a large area inaccurately and show field of view (see figure below showing a suitable technique for achieving this).

9.

Put labels outside of diagram.

10.

Label lines must point accurately to part concerned and should never cross over each other.

11.

Use minimal colour and shading.

12.

For repeating structures draw only one or two units accurately and indicate (e.g. by dotted lines), the extent of the repeating structures.

13.

Don’t draw anything until you have compared all available information (slides, models, etc.).

14.

Try to develop a 3D concept for 2D images and also try the reverse process.

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Field of veiw x 40

Lumen of duct

organ

Field of view x 100

stratified epithelium

wall of duct

Field of view x 400

ciliated layer with mucus

stratified epithelium

Figure 1: Technique to show increasing level of detail for microscope slides

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BMS192 Practical Manual

Faculty of Science Laboratories and specialist teaching spaces student induction form ______________________________________________________________________________

The Faculty of Science is committed to the safety and well being of the student population. The following are general Faculty of Science Laboratory and Specialist Teaching Spaces rules: •

Treat the facility and equipment with respect.

•

Undergraduate students are not permitted to enter or remain in the laboratories or specialist teaching spaces unless they are being supervised by a CSU staff member (unless otherwise notified).

•

Follow all safety instructions for any given practical session or exercise, including specific instructions regarding potentially hazardous material, equipment or procedures and handling human body products. All signs, documentation and Standard Operating Procedures relating to specific activities must be adhered to.

•

Bags and personal belongings should be placed in the designated area - never on the bench. Only items required should be on the bench top work area.

•

Mobile phones and music playing devices (e.g. MP3 players) must be switched off and not used. Phones used during class time may be confiscated or the user asked to leave (please see your supervisor if the phone is needed for a personal issue).

•

No smoking, drinking, eating, handling of food or application of cosmetics is permitted.

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Long hair is to be tied back at all times.

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Report all accidents, defective equipment, spillages and breakages immediately. Immediately wash skin areas that come into contact with chemicals in the laboratory.

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Under no circumstances is any unauthorised work to be conducted in the facilities.

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Always wash your hands before leaving the facilities.

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Children are not allowed in laboratories or specialist teaching spaces unless the Facility/Laboratory Manager gives prior approval for bona-fide teaching sessions for minors.

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If you have any questions regarding facility safety, please ASK your supervisor or a member of the technical staff.

•

If you have any medical (e.g. allergies) or other conditions that have the potential to impact your studies in the laboratories or specialist teaching spaces please notify your supervisor or a member of the technical staff.

•

Work areas are to be left clean and tidy. Waste must be disposed of as directed. Failure to leave work spaces in an approved manner may preclude you from subsequent classes.

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BMS192 Practical Manual

Personal Protective Equipment (PPE) A minimum standard of personal protective equipment (PPE) that complies with AS/NZS 2243.1, AS/NZS 2210.3, AS/NZS 2243.5, AS/NZS 2161, AS/NZS 1175, AS/NZS 1716 and AS/NZS 1801 will be followed in the laboratories and specialist teaching spaces. At all times PPE must be used according to standard operating procedures and instructions provided by academic and technical staff.

Required PPE may include one, many or all of the following: •

Protective clothing – properly fastened laboratory coats or disposable gowns are to be worn when required and be removed before leaving the facilities. Coats and gowns taken from the laboratory must be contained in a safe manner (e.g. by placing in sealed plastic bag). Appropriate fieldwork clothing, for example: • • • •

High Vis vests for vineyard work. Long sleeve shirts and trousers for fieldwork conducted by the School of Agricultural and Wine Science students. Overalls for fieldwork conducted by the School of Animal and Veterinary Science students. Fully enclosed footwear for fieldwork conducted by all students.

•

Eye and face protection - are to be worn when required and be removed before leaving the facilities.

•

Gloves – are to be worn when required and be removed before leaving the facilities.

•

Safety footwear – AS/NZS 2210.3 compliant (see diagram 1), closed-in, non-slip and non- permeable footwear must be worn at all times (unless notified otherwise). Shoes made of absorbent material or woven fabric (e.g. most types of runners, joggers and tennis shoes) should not be wor...