Services
In BALM our vision is to provide a state of the art imaging facility pioneering novel techniques and making cutting edge discoveries. To ensure this can be realised we have set up the facility to allow for multi-user access and polyvalent functionality of the equipment. We do our best to adapt systems to accommodate everybody's experiments and are open for users to bring in custom adaptations.
- All experiments carried out in the BALM facility are important. To ensure that you, the user have secure access to the microscopes we provide personal logins and private user space. This means you have your own settings which are modified only by you personally.
- To facilitate and ease the use of the facility for everyone we have user guidelines. Before using the facility please familarise yourself with them.
- Microscopes are expensive and fragile pieces of equipment. To ensure we keep our equipment in cutting edge condition we have to charge a small fee for usage. Our tariffs can be found on the BALM site homepage.
- We have a variety of techniques available and are always keen to develop more. If you have an idea for a project please get in touch.
- We are aiming to provide protocols and user guides for the more commonly used techniques here.
- We have a small shop of fluorescent dyes and antibodies which you are very welcome to purchase to test if your experimental conditions work. The list of what is available is here.
- Answers to the more frequently asked questions are in our FAQ section.
BALM has policies and guidelines for the imaging facility to help everyone get the best out of their experience here.
To continue the good work of the BALM facility all users must adhere to these guidelines, which have been agreed by the imaging steering committee.
Anyone who either has an interest in the work of the Imaging facility or uses the imaging facility microscopes should register with BALM by contacting the facility manager. This will allow you to be updated with news about the facility and developments in light microscopy. All users must supply an active grant code prior to facility usage.
Usage of any facility microscope is only allowed after personal training coordinated only through the imaging facility - no matter how experienced you are. This includes not just technical training, but also an introduction into facility rules, health & safety and other important issues, and therefore is compulsory. You are liable for any costs associated with damage or repair to equipment if you use BALM equipment untrained and may be barred from use of the facility.
Standard Operating Procedures for use of the machines are printed and posted above each microscope, these must be adhered to. They were written to ensure everyone has an equal opportunity to benefit fully from the equipment
All uses of BALM Microscopes must be booked on the Quartzy booking site. Please respect each other's bookings. Once booked, a person has the right to use the microscope uninterrupted. When cancelling a booking, please make every effort to contact the people booked before and after you. They might be happy to take over your time slot. – In extraordinary circumstances the BALM facility staff reserve the right to cancel bookings at short notice.
If you arrive more than 30 minutes late for a microscope session your booking is void. You will be charged for 1 hour or for the amount of time you have booked for the equipment, whichever is greater. If you cancel a session without using the equipment after the session has finished you will be liable for the cost of the whole imaging session unless you have discussed this with the BALM staff. This is to prevent people block booking busy equipment, not turning up and preventing others from using it.
Please discuss block bookings of equipment of longer than 3 hours a day with the BALM manager at the beginning of the project. The facility has over 200 users at present and everyone's experiments are equally important.
Please keep the facility safe, clean and tidy. There are clinical waste bins in every room, please use them.
Each user has an allocated user area as well as a personal profile. It is up to you to make sure there is enough space in your user area for your experiment. Please don't stop other people from doing experiments by leaving large quantities of your data on imaging facility computers.
The imaging facility cannot store your data long term. Please save your data in the location given at training. Data found on the C drive of computers, or older than 1 week on any computer attached to a microscope may be deleted if space is needed. Data on the networked mass storage server will be kept for a minimum of 1 month. After this, it will be overwritten with new data.
BALM staff are happy to advise and collaborate with projects, technical time is charged at £30 per hour. Research students and Postdocs are advised to discuss any technical work needing to be done with their PIs prior to starting a project. If there is going to be substantial technical work carried out this needs to be discussed in a meeting with both the PI and the BALM facility manager present, (at a minimum) prior to starting work.
It isn't possible for you to install personal software on any imaging facility machines.
Here is a selection of very useful links for anyone interested in microscopy:
If you've just started microscopyread this guide [PDF 1KB]about how to keep the microscope clean and working well.
Instructions about what to put in Materials and Methods about core facility microscopes are here [PDF 1KB]. You can find out more about the equipment on the equipment pages.
- Just starting out? This page tells you everything you wanted to know about how microscopy works from first principles www.microscopyu.com/articles/formulas/
- Free software that is compatible with our systems
Zen LE - www.zeiss.de/c12567be0045acf1/Contents-Frame/3f3821b370efc91cc125734c002fb38c
Imaris - http://www.bitplane.com/go/download/free-sceneviewer
Leica Lite - ftp.llt.de/softlib/LCSLite/
Image J - - rsbweb.nih.gov/ij/
FiJi - http://fiji.sc/wiki/index.php/Fiji - Am I using the correct fluorophore for my experiment? Find out here before you start. Bear in mind the laser lines on our confocals are 405, 458, 477, 488, 514, 543 and 647nm.
www.invitrogen.com/site/us/en/home/support/Research-Tools/Fluorescence-SpectraViewer.html - The Royal Microscopical Society and European Light Microscopy Initiative - Microscopy in the UK and Europe www.rms.org.uk/about-rms.shtml http://www.embl.org/elmi/
- Training courses
www.rms.org.uk/careers-courses.shtml
www.embo.org/events/calendar/practical-courses.html
www.mbl.edu/education/courses/special_topics/index.html
www.febs.org/index.php?id=86
| This list of techiques is not exhaustive. Please come and discuss with us if you want to do an experiment not listed here. We are happy to develop new methodologies or refer you elsewhere in the university. |
||
| Technique |
Microscope |
Brief overview and link |
| Epifluorescent Microscopy |
Fluorescence microscopy is a simple way of seeing where your protein of choice is located in your sample. More info here l |
|
| Confocal microscopy |
Confocal Zeiss 710 Z2, Confocal 510 Meta inverted, Spinning disk |
Confocal microscopy offers several advantages including a shallow depth of field, removal of out-of-focus glare, and the ability to collect serial optical sections from thick specimens. More info here |
| Image Analysis | See Image analysis page | Making measurements of your data for statistical analysis. |
| Colour imaging of cells |
Leica - Epi, Sterology Microscope |
To image histology and pathological specimens More info here |
| Differential Interference contrast Microscopy (DIC) |
|
DIC is a mechanism for rendering contrast in transparent specimens. More info here |
| Live cell imaging |
These microscopes will allow you to find out what’s happening in real time on your microscope |
|
| Time-lapse microscopy long term |
This microscope has a large environmental control chamber specifically for experiments lasting over 3 hours |
|
| 3 dimensional cell imaging and reconstruction |
Confocal 510 Meta inverted, Confocal Zeiss 710 Z2, Spinning disk, Imaris |
Using a automatically motorised z stage its possible to images sections of your sample and reconstruct it in three dimensions |
| Real time calcium imaging |
To quantify in real time how calcium signaling is changing |
|
| Small scale screening |
TES, Spinning Disk |
An automatic x,y,z stage makes it possible to visit each well of a 96 well dish and image it. |
| FRAP -Fluorescence recover after photobleaching |
Confocal 510 Meta inverted, Confocal Zeiss 710 Z2, Spinning disk |
FRAP measures the ability of a molecule to move around over time. Find out more here |
| FRET - Forster resonance energy transfer |
Confocal 510 Meta inverted, Confocal Zeiss 710 Z2, Spinning disk |
FRET is a mechanism describing energy transfer between two chromophores. More info here |
| Stereology |
Stereology is a standard methodology for quantitative histology used in biomedical research. Stereology.info provides a good, comprehensive overview of unbiased stereology. |
|
| Laser ablation microdissection |
Laser ablation microdissection allows the removal of very small sections of tissues or cells |
|
Below are a list of useful protocols for microscopy. This page is continuously updated so do check back.
Sample Prep
- Abcam webinar introduction to immunohistochemistry
- Although we are happy to have a go with any cell type, antibody or dye if you would like recommendations for microscopy consumables we have a list here (coming soon)
- A very general guide to immunofluorescence [PDF 1KB]
- Abcam guide to understanding secondary antibodies
- Abcam Immunohistochemistry application guide(very thorough manual for immunofluorescence) [PDF 1KB]
- Multiwell plates for fluorescent microscopy [PDF 1KB]
- Dyes for histological preparation of cells [PDF 1KB]
- Preparation of parafin embedded and frozen sections for immunofluorescence (coming soon)
- Gluteraldehyde fixation of cells [PDF 1KB]
- Methanol fixation of cells [PDF 1KB]
- Squeaky clean coverslips for perfect imaging. [PDF 1KB]
- Imaging and microscopy
- Image analysis and figures
Protocols: Microscopy
- Introduction to microscopy talk 2013 [PDF 1KB]
- Myscope online tutorials about microscopy
- Nikon Micorscopy U, all the technical facts about microscopy
- Confocal Microscopy talk 2013 [PDF 1KB]
- Long term timelapse imaging [PDF 1KB]
- Calcium imaging [PDF 1KB]
- FRET (Acceptor photobleaching) [PDF 1KB]
- FRAP [PDF 1KB]
- Laser Capture microdissection RNA [PDF 1KB] and DNA [PDF 1KB]
- Powerpoint talk about principles of confocal imaging [PDF 1KB]
- How to clean a microscope [PDF 1KB] for perfect imaging
- Using the Leica Epi to check colocalisation by eye [PDF 1KB]
Protocols: Image Analysis
Once you have acquired your images there is still plenty which can be done with them. Images can be used to make measurements or really draw out the point you need to make.
BALM offers tutorials on image analysis. You can watch these here:
- Introduction to Image analysis talk 2013 [PDF 1KB]
- Quantifying your data in the BALM facility talk 2013 [PDF 1KB]
- Metadata required for publications or MSc or PhD Theses [PDF 1KB]
- Presenting data and making figures BALM facility talk 2014 [PDF 1KB]
- Counting objects and cells [PDF 1KB]
- Measuring objects and cells [PDF 1KB]
- Counting nuclei [PDF 1KB]
- Cell morphology analysis [PDF 1KB]
- Imaris: [PDF 1KB] Image analysis for 3D images
- Imaris Learning centre - short video tutorials on Imaris
- Confocal image processing in Zen [PDF 1KB]
- The ImageJ user guide
- We use Metamorph for image acquisition and analysis. To carry out basic functions in metamorph (including how to rescale your data to 8 bit so it appears in powerpoint) seethis guide [PDF 1KB]
- A guide summarising how to use Metamorph [PDF 1KB]
- Logging data to Excel in Metamorph [PDF 1KB]
- Adding a scale bar to your image [PDF 1KB]
- Putting colour back into greyscale images [PDF 1KB]
- A
showing how to add plugins into Fiji/Image J if you aren't sure how to do this.
- Changing the bit depth of images and from 16 bit to RGB [PDF 1KB]
- Thresholding your data [PDF 1KB]
- Background subtraction [PDF 1KB]
- Making stacks and montages [PDF 1KB]
- Splitting and merging channels [PDF 1KB]
- Resizing or cropping your images [PDF 1KB]
- Colocalisation analysis [PDF 1KB]
- Using linescans to measure protein distribution [PDF 1KB]
- How to generate a scientific figure or present your data in your thesis [PDF 1KB]