- FILM-Users - imperial.ac.uk

[FILM-Users 00513] FILM Microscopy Day 2014 - user presentations needed!
by Martin Spitaler 17 Jun '14

17 Jun '14

Dear Microscopists, for the Microscopy Match Maker session in our Microscopy Day next week, we only have one presentation so far! Everyone really liked this session in previous years, but it only works if people are actually presenting - 2 slides is all that's needed, and it's all about projects and techniques to get people to know each other, you most definitely do NOT need any results (there wouldn't be enough time to show any anyway). So please recycle the last lab meeting and show us what you do. And supervisors, please encourage your students and postdoc to participate. And don't forget to register by email to Ruth O'Donnell <r.odonnell(a)imperial.ac.uk> if you plan to participate, we need the numbers for catering purposes. On that line, anyone who wants to move on later to my leaving drinks, the venue is booked now, it's the Hereford Arms <http://www.herefordarms.co.uk/>, 127 Gloucester Rd, SW7 4TE. We should be there from around 6.30pm. See you next Wednesday (with your presentation!), good imaging, Martin ------------------------------------------------------------------------ Dear microscopists, just a reminder about this year's Microscopy Day: *FILM MICROSCOPY DAY 2014, Wednesday 25 June, SAFB-G34 (South Kensington Campus), 10.00-17.00* Again a whole day full of exciting talks and presentations about microscopy, better and bigger than ever - this time with not one, but two plenary lectures, one by internal microscopist Cristina Lo Celso, the other by Phillip Kukura from Oxford. See full agenda below. And we hope many of you will add to the exciting programme with your own 3-min presenation of the microscopy you are doing in your lab: The Microscopy Matchmaker Session has always been a highlight of the day, and the more presentations we get, the more fun it will be, and the better your chances of getting to know other people's work and starting new collaborations. See also the full invitation at <https://workspace.imperial.ac.uk/imagingfacility/Public/FILM%20Microscopy%2…> Participation is free, but for catering purposes please register replying by email to Ruth O'Donnell <r.odonnell(a)imperial.ac.uk> Martin ------------------------------------------------------------------------ http://www3.imperial.ac.uk/imagingfacility/news#md _*FILM Microscopy Day - Agenda *_ *_Morning Session_*** *10.00-10.15Welcome and latest FILM News (Tony Magee) *New microscopes, new equipment, new staff *10.15-11.15 Basics of microscopy (Martin Spitaler, FILM) *Image formation, fluorescence, visualisation, quantitation */11.15-11.35 coffee & tea break/*** *11.35-12.15 Sample preparation for live and fixed samples (Stephen Rothery, FILM) *Fixatives, strategies, viability, autofluorescence ** *_Microscopy Match-Maker_*** *12.15-13.00 Microscopy Match-Maker *Research performed in FILM, presented by users through 3 min snapshot presentations */13.00-14.00 Sandwich lunch /*/with opportunity to see the new super-resolution microscope / *_Afternoon Session_*** *14.00-14.40 Super-resolution microscopy (Debora Keller, FILM) *PALM, STORM, SIM, resolution limits, artefacts */14.40-15.00 coffee & tea break/*** *_PLENARY SESSION _*** *15.00-15.55Cristina Lo Celso, Imperial College: /"Intra-vital imaging in stem cell biology"/* *15.55-16.50 Philipp Kukura, Oxford/: "Molecular Imaging"/* *16.50-17.00 Tony Magee: Round-up * ** *FILM Microscopy Day 2014: Plenary session*** *Cristina Lo Celso, Imperial College*: */"Intra-vital imaging in stem cell biology"/* After a PhD from University College London in the field of epidermal stem cell biology, Dr Lo Celso worked as a postdoc at Harvard University developing intra-vital microscopy of the haematopoietic stem cell niche. In 2009 she was appointed assistant professor at Imperial College London, where she has established a research group focussing on understanding the cellular mechanisms regulating haematopoietic stem cell (HSC) function and leukaemia development and . In close collaboration with FILM, her group has developed tools to study HSCs in their native bone marrow environment. She will present an interdisciplinary approach combining intra-vital microscopy techniques, computational image analysis, molecular profiling and mathematical modelling and the group's most recent observations of the dynamic interactions between HSCs and the bone marrow microenvironment. *Philipp Kukura, Oxford/: "Molecular Imaging" /* The primary goal of optical microscopy is to visualise microscopic structure and dynamics. Developments over the past decades have enabled routine studies down to the single molecule level and structural observations far beyond the diffraction limit through the use of fluorescence as a contrast mechanism. Despite its many advantages, one of the fundamental limitations of fluorescence detection is the frequency with which photons can be emitted and thus detected. As a consequence, imaging speeds remain limited to few to tens of frames per resulting in a considerable gap between the speed at which dynamics can be recorded and the speed of motion on the nanoscale. I will describe an alternative approach to optical microscopy that relies on the ultra-efficient detection of light scattering called interferometric scattering microscopy (iSCAT). I will show that iSCAT can follow the motion of nanoscopic labels with nm precision down to the microsecond regime, the relevant timescale for a majority of dynamics on the nanoscale. Thereby, we are able to address a surprising variety of fundamental questions ranging from the mechanism underlying processive molecular motion to the existence and formation of lipid rafts in membranes. Finally, I will show that iSCAT can detect, image and track the motion of single, proteins using light scattering without the need for any labels. _* *_ -- ------------------------------------------------------------------------ Please note that I'm leaving Imperial College on 25 June 2014. For requests after that date please contact film-service(a)imperial.ac.uk <mailto:film-service@imperial.ac.uk> *Martin Spitaler, PhD* *FILM - Facility for Imaging by Light Microscopy* - Facility Manager - Sir Alexander Fleming Building, desk 401 Imperial College London / South Kensington Exhibition Road London SW7 2AZ UK Tel. +44-(0)20-759-42023 E-mail m.spitaler(a)imperial.ac.uk <mailto:m.spitaler@imperial.ac.uk> Website: http://imperial.ac.uk/imagingfacility

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[FILM-Users 00512] Re: LacO Array
by Martin Spitaler 16 Jun '14

16 Jun '14

...forwarded on behalf of Michael - can anybody help? Best regards, Martin On 16/06/2014 14:58, Clarke, Michael wrote: > Hi Martin, > > i was wondering if you could email around and ask if anybody has a > plasmid containing an array of 128-256 LacO repeats? > > Many thanks > Michael

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[FILM-Users 00511] FILM Microscopy Day 2014
by Martin Spitaler 12 Jun '14

12 Jun '14

Dear microscopists, just a reminder about this year's Microscopy Day: *FILM MICROSCOPY DAY 2014, Wednesday 25 June, SAFB-G34 (South Kensington Campus), 10.00-17.00* Again a whole day full of exciting talks and presentations about microscopy, better and bigger than ever - this time with not one, but two plenary lectures, one by internal microscopist Cristina Lo Celso, the other by Phillip Kukura from Oxford. See full agenda below. And we hope many of you will add to the exciting programme with your own 3-min presenation of the microscopy you are doing in your lab: The Microscopy Matchmaker Session has always been a highlight of the day, and the more presentations we get, the more fun it will be, and the better your chances of getting to know other people's work and starting new collaborations. See also the full invitation at <https://workspace.imperial.ac.uk/imagingfacility/Public/FILM%20Microscopy%2…> Participation is free, but for catering purposes please register replying by email to Ruth O'Donnell <r.odonnell(a)imperial.ac.uk> Martin PS: as most of you will know by now, the Microscopy Day will also be my last day in this job before I move on, so these will also be my leaving drinks; more hardy thinkers or drinkers will probably move on to a pub nearby, tbc PPS: I know people are traditionally keen on leaving presents; to make your life easier, I propose that - unless you have an idea for something small and symbolic (easy to take on a plane) - you donate any collected money to the homeless charity <http://www.mungosbroadway.org.uk> - they are doing a really good job (and with Imperial's staff policy, you never know when you need them...) ------------------------------------------------------------------------ http://www3.imperial.ac.uk/imagingfacility/news#md _*FILM Microscopy Day - Agenda *_ *_Morning Session_*** *10.00-10.15Welcome and latest FILM News (Tony Magee) *New microscopes, new equipment, new staff *10.15-11.15 Basics of microscopy (Martin Spitaler, FILM) *Image formation, fluorescence, visualisation, quantitation */11.15-11.35 coffee & tea break/*** *11.35-12.15 Sample preparation for live and fixed samples (Stephen Rothery, FILM) *Fixatives, strategies, viability, autofluorescence ** *_Microscopy Match-Maker_*** *12.15-13.00 Microscopy Match-Maker *Research performed in FILM, presented by users through 3 min snapshot presentations */13.00-14.00 Sandwich lunch /*/with opportunity to see the new super-resolution microscope / *_Afternoon Session_*** *14.00-14.40 Super-resolution microscopy (Debora Keller, FILM) *PALM, STORM, SIM, resolution limits, artefacts */14.40-15.00 coffee & tea break/*** *_PLENARY SESSION _*** *15.00-15.55Cristina Lo Celso, Imperial College: /"Intra-vital imaging in stem cell biology"/* *15.55-16.50 Philipp Kukura, Oxford/: "Molecular Imaging"/* *16.50-17.00 Tony Magee: Round-up * ** *FILM Microscopy Day 2014: Plenary session*** *Cristina Lo Celso, Imperial College*: */"Intra-vital imaging in stem cell biology"/* After a PhD from University College London in the field of epidermal stem cell biology, Dr Lo Celso worked as a postdoc at Harvard University developing intra-vital microscopy of the haematopoietic stem cell niche. In 2009 she was appointed assistant professor at Imperial College London, where she has established a research group focussing on understanding the cellular mechanisms regulating haematopoietic stem cell (HSC) function and leukaemia development and . In close collaboration with FILM, her group has developed tools to study HSCs in their native bone marrow environment. She will present an interdisciplinary approach combining intra-vital microscopy techniques, computational image analysis, molecular profiling and mathematical modelling and the group's most recent observations of the dynamic interactions between HSCs and the bone marrow microenvironment. *Philipp Kukura, Oxford/: "Molecular Imaging" /* The primary goal of optical microscopy is to visualise microscopic structure and dynamics. Developments over the past decades have enabled routine studies down to the single molecule level and structural observations far beyond the diffraction limit through the use of fluorescence as a contrast mechanism. Despite its many advantages, one of the fundamental limitations of fluorescence detection is the frequency with which photons can be emitted and thus detected. As a consequence, imaging speeds remain limited to few to tens of frames per resulting in a considerable gap between the speed at which dynamics can be recorded and the speed of motion on the nanoscale. I will describe an alternative approach to optical microscopy that relies on the ultra-efficient detection of light scattering called interferometric scattering microscopy (iSCAT). I will show that iSCAT can follow the motion of nanoscopic labels with nm precision down to the microsecond regime, the relevant timescale for a majority of dynamics on the nanoscale. Thereby, we are able to address a surprising variety of fundamental questions ranging from the mechanism underlying processive molecular motion to the existence and formation of lipid rafts in membranes. Finally, I will show that iSCAT can detect, image and track the motion of single, proteins using light scattering without the need for any labels. _* *_ -- ------------------------------------------------------------------------ *Martin Spitaler, PhD* *FILM - Facility for Imaging by Light Microscopy* - Facility Manager - Sir Alexander Fleming Building 401 Imperial College London / South Kensington Exhibition Road London SW7 2AZ UK

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[FILM-Users 00510] Fwd: Seminar/Interview clash 20/06/2014: Ad hoc Life Sciences seminar
by Martin Spitaler 10 Jun '14

10 Jun '14

...forwarded for your information.... Martin > *From: *DoLSNEWS <dolsnews(a)imperial.ac.uk > <mailto:dolsnews@imperial.ac.uk>> > *Subject: **20/06/2014: Ad hoc Life Sciences seminar* > *Date: *June 9, 2014 4:19:07 PM GMT+01:00 > *To: *DoLS Staff and PGR Students <ls_all_dl(a)imperial.ac.uk > <mailto:ls_all_dl@imperial.ac.uk>> > > > Department of Life Sciences ad hoc seminar > > Speaker: Professor Grant J. Jensen, California Institute of > Technology, USA > > Title: "How electron cryo-tomography is opening a new window on > bacterial cell biology: type VI secretion systems and correlated light > and electron microscopies" > > Date: 12:30pm, Friday 20th June 2014, G47A Flowers Building > > Host: Dr Morgan Beeby >

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[FILM-Users 00509] Fwd: Spare inverted microscope frame
by Martin Spitaler 06 Jun '14

06 Jun '14

...forwarded for your information, Martin -------- Original Message -------- Subject: Re: Spare inverted microscope frame Date: Tue, 3 Jun 2014 10:05:06 +0100 From: Clegg, James <j.clegg11(a)imperial.ac.uk> To: Spitaler, Martin <m.spitaler(a)imperial.ac.uk> > Hi Martin > > I'm James, one of Mark Neil's students in the Physics department. We > met once about a year ago when I came round to SAF to see your old > flat panel monitors, and I've seen you at quite a few London > SuperResolution meetings. > > Mark and I are trying to build a new microscope with our own confocal > scanning apparatus but, in the true university style, we don't have a > frame to use and we probably can't afford to buy one! Perhaps this is > a bit of a long shot but Mark mentioned that you might have a frame > that we could borrow. Something like an Olympus IX71 would be ideal > as we have a few Olympus objectives already and inverted would be > best. Can you help out? > > Best regards, > > James Clegg

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[FILM-Users 00508] Fwd: CONFOCALMICROSCOPY Index - 31 May 2014 to 1 Jun 2014 (#2014-126)
by Martin Spitaler 06 Jun '14

06 Jun '14

...forwarded for your information; this should be interesting to many of you, Martin -------- Original Message -------- Subject: CONFOCALMICROSCOPY Index - 31 May 2014 to 1 Jun 2014 (#2014-126) Date: Mon, 2 Jun 2014 00:03:15 -0500 From: CONFOCALMICROSCOPY automatic digest system <LISTSERV(a)LISTS.UMN.EDU> Reply-To: <CONFOCALMICROSCOPY-Search-request(a)LISTS.UMN.EDU> To: <CONFOCALMICROSCOPY(a)LISTS.UMN.EDU> CONFOCALMICROSCOPY Index - 31 May 2014 to 1 Jun 2014 (#2014-126) LISTSERV mailing list manager <http://www.lsoft.com> LISTSERV 15.0 <http://lists.umn.edu/cgi-bin/wa?LIST=CONFOCALMICROSCOPY> CONFOCALMICROSCOPY Index - 31 May 2014 to 1 Jun 2014 (#2014-126) Index Date Lines Subject and Sender 043991 <http://lists.umn.edu/cgi-bin/wa?A2=ind1406&L=CONFOCALMICROSCOPY&P=R2> 06/01 52 EMBO practical course on Biomolecular interaction analysis: From molecules to cells From: Paula Sampaio <sampaio(a)IBMC.UP.PT> Browse the CONFOCALMICROSCOPY online archives. <http://lists.umn.edu/cgi-bin/wa?LIST=CONFOCALMICROSCOPY> Powered by the LISTSERV Email List Manager <http://www.lsoft.com/products/listserv-powered.asp>

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[FILM-Users 00507] Fwd: Bus survey - June 2014
by Martin Spitaler 05 Jun '14

05 Jun '14

...forwarded for your information: All FILM Users commuting between FILM branches in Hammersmith and South Kensington should be interested, Martin -------- Original Message -------- Subject: Bus survey - June 2014 Date: Mon, 2 Jun 2014 12:59:54 +0100 From: Faculty of Medicine <medicine-notifications(a)imperial.ac.uk> To: <m.spitaler(a)imperial.ac.uk> *Survey: Study on potential Bus Service, Hammersmith Campus to South Kensington Campus * /Sent on behalf of the Chief Operations Officer to all Faculty of Medicine staff/ Dear Colleague, As announced in the last Staff Briefing (22 May), the College is conducting a study to establish the current level of demand for direct transport between the South Kensington Campus and the Hammersmith Campus (the Hammersmith Hospital, Du Cane Road), with a view to potentially running a direct College bus service between the two campuses. In order to establish the current level of demand for such a service, we would appreciate your input via the survey <https://www.surveymonkey.com/s/2Y2YLMC>. *Deadline for completion is Friday 6 June 2014*. Should demand be sufficient, a pilot service will be set up. The survey should only take a few minutes to complete, and there is a £100 Amazon voucher to be won. Thank you in advance for your help. Best wishes, *Simon Harding-Roots *Chief Operations Officer* * *Survey link: *https://www.surveymonkey.com/s/2Y2YLMC ------------------------------------------------------------------------ view this email online <https://fomemailer.imperial.ac.uk/resource/email/d07792c0-9860-44ed-bcec-91…>

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[FILM-Users 00506] FILM Microscopy Day 2014
by Martin Spitaler 31 May '14

31 May '14

Dear microscopists, the academic year is approaching its end, and as usual we conclude our FILM Club series with the FILM Microscopy Day 2014: *FILM MICROSCOPY DAY 2014, Wednesday 25 June, SAFB-G34 (South Kensington Campus), 10.00-17.00* Again a whole day full of exciting talks and presentations about microscopy, better and bigger than ever - this time with not one, but two plenary lectures, one by internal microscopist Cristina Lo Celso, the other by Phillip Kukura from Oxford. And we hope many of you will add to the exciting programme with your own 3-min presenation of the microscopy you are doing in your lab: The Microscopy Matchmaker Session has always been a highlight of the day, and the more presentations we get, the more fun it will be, and the better your chances of getting to know other people's work and starting new collaborations. See also the full invitation at <https://workspace.imperial.ac.uk/imagingfacility/Public/FILM%20Microscopy%2…> So keep the day free, good imaging, Martin http://www3.imperial.ac.uk/imagingfacility/news#md _*Agenda *_ *_Morning Session_*** *10.00-10.15Welcome and latest FILM News (Tony Magee) *New microscopes, new equipment, new staff *10.15-11.15 Basics of microscopy (Martin Spitaler, FILM) *Image formation, fluorescence, visualisation, quantitation */11.15-11.35 coffee & tea break/*** *11.35-12.15 Sample preparation for live and fixed samples (Stephen Rothery, FILM) *Fixatives, strategies, viability, autofluorescence ** *_Microscopy Match-Maker_*** *12.15-13.00 Microscopy Match-Maker *Research performed in FILM, presented by users through 3 min snapshot presentations */13.00-14.00 Sandwich lunch /*/with opportunity to see the new super-resolution microscope / *_Afternoon Session_*** *14.00-14.40 Super-resolution microscopy (Debora Keller, FILM) *PALM, STORM, SIM, resolution limits, artefacts */14.40-15.00 coffee & tea break/*** *_PLENARY SESSION _*** *15.00-15.55Cristina Lo Celso, Imperial College: /"Intra-vital imaging in stem cell biology"/* *15.55-16.50 Philipp Kukura, Oxford/: "Molecular Imaging"/* *16.50-17.00 Tony Magee: Round-up * ** *FILM Microscopy Day 2014: Plenary session*** *Cristina Lo Celso, Imperial College*: */"Intra-vital imaging in stem cell biology"/* After a PhD from University College London in the field of epidermal stem cell biology, Dr Lo Celso worked as a postdoc at Harvard University developing intra-vital microscopy of the haematopoietic stem cell niche. In 2009 she was appointed assistant professor at Imperial College London, where she has established a research group focussing on understanding the cellular mechanisms regulating haematopoietic stem cell (HSC) function and leukaemia development and . In close collaboration with FILM, her group has developed tools to study HSCs in their native bone marrow environment. She will present an interdisciplinary approach combining intra-vital microscopy techniques, computational image analysis, molecular profiling and mathematical modelling and the group's most recent observations of the dynamic interactions between HSCs and the bone marrow microenvironment. *Philipp Kukura, Oxford/: "Molecular Imaging" /* The primary goal of optical microscopy is to visualise microscopic structure and dynamics. Developments over the past decades have enabled routine studies down to the single molecule level and structural observations far beyond the diffraction limit through the use of fluorescence as a contrast mechanism. Despite its many advantages, one of the fundamental limitations of fluorescence detection is the frequency with which photons can be emitted and thus detected. As a consequence, imaging speeds remain limited to few to tens of frames per resulting in a considerable gap between the speed at which dynamics can be recorded and the speed of motion on the nanoscale. I will describe an alternative approach to optical microscopy that relies on the ultra-efficient detection of light scattering called interferometric scattering microscopy (iSCAT). I will show that iSCAT can follow the motion of nanoscopic labels with nm precision down to the microsecond regime, the relevant timescale for a majority of dynamics on the nanoscale. Thereby, we are able to address a surprising variety of fundamental questions ranging from the mechanism underlying processive molecular motion to the existence and formation of lipid rafts in membranes. Finally, I will show that iSCAT can detect, image and track the motion of single, proteins using light scattering without the need for any labels. _* *_ -- ------------------------------------------------------------------------ *Martin Spitaler, PhD* *FILM - Facility for Imaging by Light Microscopy* - Facility Manager - Sir Alexander Fleming Building 401 Imperial College London / South Kensington Exhibition Road London SW7 2AZ UK

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[FILM-Users 00504] looking after the microscopes
by Martin Spitaler 29 May '14

29 May '14

Dear microscopists, today unfortunately I have to remind you to be more careful and cooperative looking after FILM microscopes - some of you have become quite reckless, and this is creating lost time, bad image quality and other problems for other users: * Please be _*very*_ _*careful with the lenses*_: Someone has just damaged the 63x water immersion objective on Confocal 3, we have to send it in for repair, which will probably cost at least £1,000; if you can't find the focus, try with low magnification first, and go to the edge of your slide, dish or coverslip, which is always easy to see. * When using oil immersion objective lenses, only use a _*small drop of oil*_, and wipe the lens between samples. Recently, microscopes were regularly soaked with immersion oil (in particular on CF4), which went into the microscope body, onto all air objectives and everywhere else. This means that users get bad quality images, and staff spend a long time cleaning microscope and lenses. * The *stage insert for multiwell plates *for Leica confocals had disappeared and was urgently needed for a project; we had to order a replacement, and suddenly it has reappeared - another £450 wasted. * _*Rubbish:*_ Please clean the microscope space after yourself, put all rubbish in the appropriate bin: Lab waste into clinical waste bins, everything else into the normal bins - do _*not*_ through any coffee cups or similar into the clinical waste bins, it costs a lot of money to have them incinerated! * If you spill liquid or break glass, please also clean it up! It's not acceptable if the next microscope user has to spend the first part of the session to clean the space (as happened recently on CF3 with broken glass). If things go wrong and you don't know how best to clean it, please ask. I hope this will help to remind the few people behaving badly what effect their negligence has and do better in the future. Thanks for your cooperation, good imaging, Martin -- ------------------------------------------------------------------------ *Martin Spitaler, PhD* *FILM - Facility for Imaging by Light Microscopy* - Facility Manager - Sir Alexander Fleming Building, desk 401 Imperial College London / South Kensington Exhibition Road London SW7 2AZ UK Tel. +44-(0)20-759-42023 E-mail m.spitaler(a)imperial.ac.uk <mailto:m.spitaler@imperial.ac.uk> Website: http://imperial.ac.uk/imagingfacility

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[FILM-Users 00503] new ONIX perfusion system available in FILM
by Martin Spitaler 28 May '14

28 May '14

Dear microscopists, thanks to a grant application organised mainly by Brian Robertson, we have now a new perfusion system available in FILM, the CellAsic ONIX microfluidic platform: * http://www.cellasic.com/ONIX.html * http://www.millipore.com/life_sciences/flx4/cellasic_live_cell_imaging#tab1… (the system was recently bought by Millipore) Currently, the system is implemented on Widefield 2, but the perfusion plates are the size of a multiwell plate, so in principle fits on any inverted microscope (the software needs to be installed). Plates are available for bacteria, yeast and mammalian cells, but after the take-over by Millipore a new round of standardisation and upscaling has resulted in temporary shortages for some of the plates. Application notes and descriptions - including some impressive videos - are available on the website and also on the FILM Documentation Server (\\film-nas.med.ic.ac.uk\documentation) in the folder "...\Cellasic\". The system is already pretty busy, but if you are interested, please get in touch with FILM staff. Good imaging, Martin -- ------------------------------------------------------------------------ *Martin Spitaler, PhD* *FILM - Facility for Imaging by Light Microscopy* - Facility Manager - Sir Alexander Fleming Building, desk 401 Imperial College London / South Kensington Exhibition Road London SW7 2AZ UK Tel. +44-(0)20-759-42023 E-mail m.spitaler(a)imperial.ac.uk <mailto:m.spitaler@imperial.ac.uk> Website: http://imperial.ac.uk/imagingfacility

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