Information for users
We are using iLab for registration and booking purposes.
Please go here
Screening workflow before applying for time (assumes your have 2D class averages from negative staining for SPA):
1. Freeze 4-6 grids with your sample (macromolecule, cell or nano crystal)
same vitrobot conditions ( blot paper saturation, humidity, temperature ....)
2. Screen one of those
on a 120kV microscope (eg. L120C) for SPA, or 200-300kV (eg. TF30) for tomo and microED
- A. if it is good, acquire 1 grid atlas (or very low mag, eg. x50), at least 2 grid squarres, at least 4 grid holes
- B. if it is not good try another of you 4-6 grids
- if it is good go to A
- if it is not good try again or start from scratch
- Apply for Single Particle project
You will need 2D classes of negative staining, an atlas, low lag, high mag of a cryoEM screening grid
- Apply for tomography project
you will need an atlas of a screening grid
- Apply for MicroED project
You will need an optical microscopy image of your crystal tray, an atlas, low mag magnification and high magnification of a cryoEM screening grid
Electron microscopes charges 2020 from 18th of January 2021 | |
|---|---|
Beam time for all Instruments^ | |
| ^ not for blockface imaging and Arctica (see below) | |
| Subscription (only for The University of Melbourne research groups with Themis account string) | $ upfront |
| 50h per research group | 3890 |
| 100h per research group | 6962 |
| 200h per research group | 12285 |
| 400h per research group | 21294 |
| User Operated | $/h |
| University of Melbourne | 82 |
| Other Research Institutes | 119 |
| Corporate | 211 |
| Unit Staff Operated ($/h) | $/h |
| University of Melbourne | 177 |
| Other Research Institutes | 211 |
| Corporate | 306 |
| Training ($/h) * | $/h |
| University of Melbourne | 128 |
| Other Research Institutes | 164 |
| Corporate | 260 |
Block face imaging (VolumeScope) | |
| Sample set up and acquisiton included (minimum 18h run) | $/h |
| Standard overnight run LIEF partners | $35/h |
| Standard overnight run Non LIEF partners | $57/h |
Arctica Automated cryoEM | |
Sample set up and acquisition included with 3 grids/autogrids, 1 day= 20h regulated access ^^ | $/day |
| LIEF partners | $1460 |
| Non LIEF partners | $2260 |
Ancillary equipment: | |
| Carbon coater Emitech K950X (Loc: Bio21) | free# |
| Plasma cleaner (Loc: Bio21) | free# |
| Gold sputter coater Dynavac (Loc: Bio21) | free# |
| Dynavac coating (Loc: Botany) | free# |
| Vitrobot (Loc: Bio21 rate /1h) | free# |
| Leica AFS2 (rate/run) | from $176 |
| Leica EMPACT2 (rate/2h) | from $118 |
| cryo ultra microtome (rate /2h) | from $57 |
| ultra microtome (rate /h) | from $13 |
* Is considered "training" a session in which the aim is not too look at a sample but to teach the user how to use the instrument. The user can bring his/her own sample. After 4h it will be up to the unit staff to decide if further "training" is considered as "training" or "staff operated"
# When using microscope of the Advanced Microscopy Facility
It is the responsibility of the user to turn up on time. Charges will be made according to booking times. Cancellation have to notified at least 24h prior to booking starting time. Full charge will be incured if the notification is not received on time.
Subscription: The subscription is for each research group regardless of the number of group member. It is valid for 12 months from the date of purchase, when all the hours are used the research group will revert to the standard hourly fee. Unused hours will expire after 12 months from the date of purchase. Only valid for user operated microscope time. Valid across all instruments. subscription will commence when payment has been cleared
^^Arctica access policy: users must have shown relevant staff negative staining and corresponding class averages as well as cryoEM screening grids micrographs. Application for access: Assoc Prof Eric Hanssen
- Societies
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Web resources
MyScope, microscopy tutorial (pre-requisite for one on one teaching at IHIC)
Leica Science-Lab, tutorials, article about microscopy and applications
EM University, large repository of knowledge on EM and cryoEM
CryoEM101, videos and animation about the cryoEM workflow
- Instrument manuals
-
Calculators
Basic SPA box size calculator
Assuming 2.7 mm Cs
SPA refinement memory calculator
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What to bring
Tolls you will need:
Minimum kit:
- 2 sets of Dumont tweezers style #5, non-magnetic
- appropriate grids (see below)
- grid storage
Optional tools:
- ThermoFisher Vitrobot or Leica EMGP tweezers
- Diamond knife for ultrathing sections (35 degres for standard sections, 45 degres for hard materials) and cryo ultrathin sections (IHIC will provide glass knives only)
Recommanded grid types by application:
- negative staining and small nanoparticles: 400 mesh carbon coated grids (copper)
- plastic sections; 50-100 mesh formvar carbon coated grids (copper)
- plastic section tomography: 200 mesh formavar carbon coated grids (copper)
- immunolabelling: 100-200 mesh formvar carbon coated grids (gold, nickel or copper)
- routine cryoEM (liposome/exosome/cubosomes/phages/bacteria ...): 300 mesh lacey carbon (copper)
- single particle analysis: 1.2/1.3 (or smaller holes) UltraFoil (or equivalent).
Publication network 2010-2019 Bio21 AMF
199 research groups
541 publications in peer reviewed journals
173 co-authorship by Bio21 AMF staff members
Thank you to Mr Roger Curtain and Mr Zlatan Trifunovic who combed through over 20,000 publications.
Please acknowledge us in your publications to make the next iteration easier.
Click on the map for more details (java needs to be installed)
