TITLE OF MODULE: Skills - Cell Imaging
MODULE NUMBER: BIO00032I
ORGANISER: Gonzalo Blanco
SUBJECT COMMITTEE: COB
VERSION: May 2014
TERM TAUGHT: Spring 2015
PREREQUISITES: It is recommended that module BIO00035I, Cell biology, is taken
alongside this module.
ASSESSMENT: Two separate open assessments for each half of the module, each
1000 words approximately and comprising a series of questions requiring short
answers of between 20-200 words. The assessments will require submission of the
completed script by the end of week 10 of the Spring term.
Mod Code
Mod Name
Level
Credits
Owner
Module type
Compensatable
Reassessable
BIO00032I
Skills – Cell Imaging
Intermediate
5
Gonzalo Blanco, Biology
Y
N
Module availability
Occurrence
1
Terms taught
Term 5
Assessment Information
Sequence
Assessment title
Type
Weight
Final
Assessment?
001
The use of in vitro-cell culture,
transfection and imaging in cell biology
(GB)
CSWK
50%
Yes
002
Imaging neuromuscular junctions with
fluorescent proteins and antibodies
(SS)
CSWK
50%
Yes
1
SUMMARY:
Two experimental systems will be used to introduce techniques such as in vitro cell
culture, transfection, staining and fluorescent microscopy on cells and whole
organisms. The students will gain an appreciation of how different techniques can be
used to study the biology of single cells and tissues and how they can be used in
combination to answer specific questions relating to cell function.
AIMS:
This module aims to give students a theoretical and practical understanding of some
of the techniques commonly used in cell and organismal biology. The students will
receive introductory explanations of the theory behind the use of different
techniques, and will then gain direct practical experience of these approaches. Skills
acquired in this module will aid students wishing to undertake final year research
projects particularly related to animal cell biology.
LEARNING OUTCOMES:
Upon completing this module, students will have the ability to:





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Growth and maintain mammalian cells in vitro.
Use transfection to study the biology of the mammalian cell, and examine the
expression of molecules by fluorescence microscopy.
Design labelling procedures and the appropriate use of controls to confirm
specificity of the labelling procedure.
Observe and analyse qualitatively and quantitatively changes in cellular
phenotypes using fluorescence microscopy.
Use literature available in the library, the vle and via the Internet to further
explore advances in techniques relevant to the study of animal cell biology.
Acquire and interpret information from fluorescent images of a whole animal
preparation.
SYNOPSIS:
Lecture 1 (GB+SS)
Introduction
Overview of the module, including the scientific basis of the techniques that will be
taught in the practicals.
(Timetable 1 h, preferably week 2)
Practicals 1-6
The practicals cover six 2 or 3 h sessions and a final 1 h interpretative session. The
cohort will be split into two equal sized groups of 12 students, A and B.
Groups A&B will attend the Introductory lecture (week 2) and debriefing session
(week 5).
For the practical work, groups A and B should be timetabled concurrently:
Group A will do Practicals 1-3 with GB in week 3 and Practicals 4-6 with SS in week
4.
Group B will do Practicals 1-3 with SS in week 3 and Practicals 4-6 with GB in week
4.
The practical work will cover two experimental systems:
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The use of fluorescence microscopy in cell biology (GB).
The aim of this section is to teach the students different ways of studying the
subcellular localisation of proteins. Fibroblast cells will be transfected to overexpress
fluorescently tagged proteins of interest. The localisation and effect of the proteins
on cell morphology will be analysed.
Practical 1 and 4: Students will be taught the basics of tissue culture. This will
include how to work under sterile conditions, how to culture cells, prepare seeding
dilutions and seed onto coverslips.
(Timetable: 3 hours – preferably morning)
Practical 2 and 5: Students will perform transient transfection of cells seeded on
coverslips using the Ca transfection method. Mammalian expression constructs that
can be visualised by fluorescence microscopy are available. They encode proteins
fused to enhanced green fluorescent protein (GFP). This session introduces the
basics of overexpression analysis to the students.
(Timetable: 3 hours, 2 days after Practical 1)
Practical 3 and 6: Students will be taught how to stain the cells they seeded on day
1 with fluorescent markers to visualise the cytoskeleton and nuclei of COS7 cells.
Students will analyse the cells transfected in Practical 2 using fluorescence
microscopy to visualise the subcellular localisation of the overexpressed protein and
its effect upon cell morphology.
(Timetable: 3 hours, 2 days after Practical 2, and preferably in the morning)
Imaging neuromuscular junctions with fluorescent proteins and antibodies
(SS).
This part of the course will generate an image of the muscle and the synapse using
Drosophila larvae of two genotypes. The size of the muscle and the length of the
synapse will be measured for the two genotypes and compared to determine the
effect of the mutation on synapse growth.
Practical 1 and 4: Students will practice dissecting Drosophila 3rd instar larvae and
be taught how to recognise and select for male larvae. All of this analysis will be
done in males.
(Timetable: 3 hours)
Practical 2 and 5: Students will dissect and fix in formaldehyde, Drosophila larvae,
sufficient for a simple statistical analysis, they will then add an antibody to the
preparation and incubate until the next session.
(Timetable 3 hours)
Practical 3 and 6: Students will wash the preparations with saline and detergent,
and add them to a glycerol solution for a short period. The preparations will then be
mounted on microscope slides and imaged on the fluorescent microscope.
(Timetable: 2 hours, 48hrs after session 2)
MICROSCOPY SESSIONS: These sessions include an introduction to the
fluorescent microscope, which will then be used by the students to analyse and
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collect data on the subcellular localisation of the overexpressed protein and its effect
upon cell morphology and actin cytoskeleton from the slides from Practical 3 and 6
(GB), which will be used to answer a significant part of the open assessment.
Likewise, microscopy sessions will be required to image synaptic structures from the
processed dissected Drosophila larvae preparations from Practicals 3 and 6 (SS).
This session will be arranged with groups of students according to the their
academic timetable and availability of fluorescent microscopes.
Lecture 2 (GB + SS)
A debriefing session will allow students to ask questions about their practical work
and the interpretation of their data.
In addition, the assessment will be distributed at this session.
(Timetable 1h in the week after Practical 6)
RECOMMENDED READING:
Any of the following textbooks will help understanding of the course but it is not
required for students to purchase the books unless they have already done so as
part of their studies for modules in Cell Biology, Neurophysiology or Immunology.
The books might also be useful for modules in the final year including Advanced
Topics in Immunology, Cancer and Cell Cycle and Neuroscience.
 Cassunmeris, L., Lingappa, V.R., Plopper, G. (2011) Lewin’s Cells (2nd edition)
Jones and Bartlett publishers.
 Janeway, C. A. & Travers, P. (2005) Immunobiology. (6th edition) Churchill
Livingstone.
 Purves et al. (2004) Neuroscience. (3rd Edition) Sinauer Associates Inc.
LECTURERS AND ORGANISATION:
L1 GB & SS
P1-6 GB & SS
L2 GB & SS
DEMONSTRATING INFORMATION (IF APPLICABLE):
1 demonstrator with knowledge of cell biology for the duration of all GB practicals.
1 demonstrator with knowledge of fly larvae dissections for the duration of all SS
practicals.
Demonstrators will not be needed for the final debriefing session.
MAXIMUM NUMBERS: 24
STUDENT WORKLOAD:
Lectures:
2h
4
Practicals: 16 h
Total Contact hours: 18 h
Private study: 32 h
5
STAFF TEACHING COMMITMENTS: Please enter the total number of sessions attended by each staff member:
Staff initials
GB
SS
Lecture sess.
2
2
Practical sess.
6
6
Other (specify)
SAFETY AND TIMETABLING INFORMATION:
Please fill in an entry in the table below for each teaching session in the module. Where possible group sessions that
have identical entries in all columns. A key below explains the column headings.
Session
Type
Occ.
Haz.
Max.
Equipment
Lecturers
Description
Lectures 1-2
L
1
A
24
N/A
GB + SS
Introductory and debriefing sessions
1
C
12
H, S, M
GB
The use of fluorescence microscopy
in cell biology
1
C
12
H, M
SS
Imaging neuromuscular junctions
with fluorescent proteins and
antibodies
1h
Practicals 1-6
P
3h
Practicals 1-6
P
3h
KEY: Sess: session number or group of sessions (e.g. 6, 8-13). Type: Type & Duration; L 1 hr-lecture, P 3 hr-practical class, S 1 hr-seminar, T 1 hr tutorial. Specify non-standard
type or duration. Occ.: number of occurrences of the session (e.g. 3 occurrences, each taking one third of the class). Haz.: hazard rating: A, low hazard, lectures, 'paper & pencil'
problem sessions, etc; B, medium hazard, observational practicals where students move about but are not involved in C category activities; C, high hazard, practicals involving
potential hazard in overcrowded laboratories, e.g. naked flames, hot liquids, glassware, pipetting. Max.: maximum number of students permitted in session, which may be less than
the maximum number taking the module in, for example, a circus practical (see maximum room capacities above). Equipment: essential equipment in limited supply: M microscopes,
D dissecting microscopes, C computers, S spectrophotometers, Ch chart recorders, H haemocytometers, Cm microcentrifuges, Cc cooled centrifuges, Cb bench centrifuges, G Gilson
pipettes, specify other items. Lecturers: the initials of the lecturers participating in the session. Description: a brief description of the session(s).
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