Universität Wien FIND

300104 UE Hands on Cryopreparation of Biological Samples for Biological Studies (2019W)

5.00 ECTS (4.00 SWS), SPL 30 - Biologie
Continuous assessment of course work

Registration/Deregistration

Details

max. 6 participants
Language: German, English

Lecturers

Classes

Attention! No online registration possible.
The course targets students who plan cryopreparation and TEM in the EM lab of the CIUS as part of a project, as well as students interested otherwise (max. 6 participants).
Please, contact Siegfried Reipert until 31st October 2019 (siegfried.reipert@univie.ac.at; Althanstr. 14, UZAI, Raum 1.111). Potential participants and students ranked first on the waiting list will be invited via e-mail to participate in the obligatory safety instruction.

Obligatory safety instructions will be given by Daniela Gruber on 13th November 2019, 9 a.m. to 10 a.m.; Ü2, UZA 1, Biozentrum Althanstraße 14, 1090 Vienna.

Thursday. 14.11.2019 09:00 – 12:00 lecture Ü2, 13:00 – 17:00 Practical work, EM-LAB Core Facility Cell Imaging und Ultrastrukturforschung.
Friday. 15.11.2019 09:00 – 17:00 Practical work, EM-LAB Core Facility Cell Imaging und Ultrastrukturforschung.
Monday, 18.11.2019 09:00 – 17:00 Practical work, EM-LAB Core Facility Cell Imaging und Ultrastrukturforschung.
Tuesday. 19.11.2019 09:00 – 17:00 Practical work, EM-LAB Core Facility Cell Imaging und Ultrastrukturforschung.
Wednesday. 20.11.2019 09:00 – 17:00 Practical work, EM-LAB Core Facility Cell Imaging und Ultrastrukturforschung.
21st Nov. - 28th Nov 2019 Personal appointments for cutting TEM sections with an ultramicrotome. EM-LAB Core Facility Cell Imaging und Ultrastrukturforschung.
Monday, 2.12. 2019 9:00-17:00; Group A: Work with the TEM. EM-LAB Core Facility Cell Imaging und Ultrastrukturforschung.
Tuesday, 3.12. 2019 9:00-17:00; Group B: Work with the TEM. EM-LAB Core Facility Cell Imaging und Ultrastrukturforschung.
Friday, 6.12.2019, 9:00 – 13:00 Final discussion, presentation of the result (talks). Conference Room Zoology (1.015)


Information

Aims, contents and method of the course

The life-science community increasingly includes cryopreparation techniques for microscopic and analytical methods. Ultrarapid cryoimmobilization of living samples in combination with low-temperature dehydration-and fixation (freeze substitution) became a quality standard for the preservation of animal- and plant tissues for transmission electron microscopy that enabled new approaches to tackle biological questions.
The course educates students in the principles of two substantially different freezing techniques, namely, high-pressure freezing and plunge freezing. Both techniques will be used during the course to freeze selected biological samples that are either alive or chemically fixed (as they result frequently from field experiments). The samples will be freeze substituted by using agitation modules inserted in the cryochambers of automated freeze substitution systems (AFS). The application of those modules, developed in our EM lab, allows drastic reduction of the processing time (see literature). Subsequently, the samples will be embedded either in epoxy resin or Lowicryl. The latter is in particular of interest for applications that aim on immunolabeling.
The sectioning of the resin embedded samples will be arranged according to individual requirements for practicing within a separate stage of the course. Practical work will be complemented by lectures and discussions, which will highlight the role of cryopreparation in the concept of biological sciences and its combination with other methods. Finally, the sample quality will be evaluated in the light- and electron microscope. The results will be presented in a short talk.
In consequence, the course provides a preparative base for more complex experimental approaches, which may include phenotyping at ultrastructural level, labeling methods (immunolabeling, in situ hybridization), element analysis (EDX and EELS) and, increasingly, also NanoSIMS.

Assessment and permitted materials

Participation: 40%
Oral presentation of the results: 40% (23th March 2018)
Protocol: 20%
Every item has to be positive.

Minimum requirements and assessment criteria

Theoretical knowledge (lecture). Application of equipment for sample preparation and TEMs (practical work). Evaluation, discussion and interpretation of the results.

Examination topics

Ability to select suitable methods for cryopreparation for TEM samples, practical work in our chemistry lab, save handling of cryogens, liquid nitrogen and propane, handling of equipment for sample preparation incl. plunge freezer, high-pressure freezer, ultrathin sectioning with ultramicrotomes, work with TEMs, interpretation and evaluation of the results.

Reading list


Association in the course directory

MBO 7, MGE III-2, MMB W-2, MMEI III, MZO 2, MZO W-1, WZB, M-WZB

Last modified: We 25.09.2019 12:28