When: June 2nd-4th , 2015.
Where : TAC building
300 Cedar Street
(corner of Cedar and Congress Ave)
New Haven CT, USA
The afternoon symposia will take place in the TAC building auditorium (N107). The microscopy equipment will be available for use in a series of contiguous rooms on the 2nd floor directly above the auditorium (N221-N231). Some equipment in other campus buildings may also be demonstrated or available for use.
Yes, we STRONGLY ENCOURAGE you bring your own samples! Where else can you see your samples on so many different instruments in the same day?! You can bring fixed or living specimens, but these must be limited to BLS1 samples. BLS2 samples are not permitted. If you need a sample dish to be held at 37 C, 5% CO2 in a nearby lab please write to one of the organizers. Please note that there will not be an incubator in the rooms housing the microscopes.
EM samples: While EM imaging can be performed with samples we have prepared that will be provided on site, if you are interested in imaging your OWN samples, they must be prepared prior to the workshop. Most EM sample preparation requires several days of processing, and hence cannot be completed from beginning to end while at the workshop. Preparation for SEM prior to the workshop is fairly straightforward, as described in these suggested protocols. You merely need to pass your sample through the fixative solutions and then while at the workshop, sample preparation can be completed using the Leica CDP300 instrument that will be available to complete the dehydration and metal coating needed for SEM. Then take it to the JEOL Neoscope to obtain SEM images! Preparation for TEM is more complex as it requires fixation followed by treatment with hazardous chemicals over the course of several days. For those investigators at Yale, fixative solutions can be obtained from the Yale EM facility staff that will allow you to fix your samples as described in these protocols for tissues or cultured cells. The EM facility has generously offered to complete the labeling process for you, as they cannot provide the hazardous chemicals required for the remaining steps BUT preparations for this must be arranged in advance. Last minute requests may not be possible. Please contact Xinran Liu (firstname.lastname@example.org) with any further questions about EM sample preparation.
Super Resolution samples: The STED super resolution microscope available for use during the workshop has restrictions in the fluorophores that can be used. Attendees are strongly advised to follow manufacturer protocols for sample preparation. A protocol can be downloaded from the links below.
Although entrance to the rooms and viewing of the microscopy equipment does not require a reservation, hands-on access to some of the equipment can not be guaranteed without advance reservation. Most of the time, these microscopes will be available for use outside of class times, but if you would like to guarantee access to a particular instrument you should reserve time on it through our online scheduling system.
Registered individuals will receive by email a username and password that can be used to reserve equipment on the website ScheduleBook.com. After logging onto the site in the lower right hand corner of the homepage, registrants can reserve a specific piece of equipment by first choosing the preferred date by clicking on the "pick date" button in the upper right hand quadrant of the screen. Then click on the “event” button in the upper right hand quadrant of the screen. Please record your cell phone number in the comments box so that we may contact you to discuss the reservation if necessary. Reservations can be edited or deleted by clicking on the small “E” or “D” next to your name within the time slot.
Although you may reserve up to 4 different pieces of equipment, we request that reservations be kept to a minimum in order to allow as many individuals access to the equipment as possible. Out of respect for others, please delete your reservations as soon as you know that you will not need them.
Representatives from each of the microscope companies will be available to guide interested individuals in performing any technique that is applicable to the available equipment and samples. Of course you are welcome to bring any BSL1 sample with you that would suit your application of interest! The samples provided at this year's workshop will be appropriate for traditional confocal, super resolution and spinning disk microscopy, but these preparations would not be very interesting for FRET or time resolved imaging. Thick fixed specimens will be available at the LaVision BioTec two-photon LSM.
Prepared samples will be available at the scanning and transmission EM microscopes as well. If you would like to bring your own samples, please note that sample preparation for TEM and SEM requires several days of processing. Please see the section on "Can I really bring my own samples?" for a discussion of sample preparation.
Post acquisition image analysis is also available during software "walk-in clinics". Vendor representatives for Imaris will be present on Tuesday June 2nd to try out analysis of either your own data or exemplary data sets that they can provide to demonstrate 3D visualization, cell tracking and tomographic analyses.
Visit the following website for the location of the TAC building and suggested parking within the medical campus:
http://www.yale.edu/about/maps.html - For additional maps and resources
The majority of the available equipment installed by vendors will be located within contiguous teaching labs within the North TAC building on the 2nd floor. We take advantage of the existence of some specialized facilities on campus as well, for example the EM facility and an intravital imaging facility that contains a two-photon laser scanning microscope. These are located in adjacent buildings that are easily found by following prominent signs, but of course staff at the registration table will be happy to give you directions.
There are two Lightsheet systems that are available for use during the 2015 workshop, made by LaVision Biotec and Leica. The LaVision system will be installed within the TAC building. Please note that the Leica DLS Lightsheet will be installed on another distant portion of the Yale campus known as the West Campus within the Leica Center of Excellence of the WC Imaging Core. Researchers joining in a small group practical will be taken there at 10:05 AM and returned afterwards by a small private bus to that class in the morning, but if you have signed up for a session in the afternoon you will need to take the Purple route Yale shuttle system to West Campus. Here is a link to the Yale shuttle schedule, a real-time indicator of shuttle bus position, and a pdf further describing the location of the WC Imaging Core.
Light Sheet microscopes are usually employed for two very different types of applications: 1) the imaging of very thick tissue samples that have undergone "clearing" and 2) the imaging of small transparent organisms like zebrafish embryos or C. Elegans over time to observe development or rapid events.
For the preparation of small transparent organisms,please see this protocol offered by Leica. This embedding approach can also be used for the LaVision system as well. The final trimming of the agarose should take place at the microscope however, since the final size is important and can be easily done together with the vendor representative.
There are now many protocols published in the literature for the clearing of thick tissues to increase their transparency. The final step for many of these protocols is an aqueous buffer, but some use organic solvents. Leica is not accepting samples in organic solvents at this time, but such samples can be imaged with the LaVision microscope during the workshop.
Cleared tissues in aqueous buffers: Samples prepared with a final buffer that is aqueous based and not an organic solvent can be imaged at either the LaVision or Leica Lightsheet microscopes. If possible, bring more of the final buffer with you to serve as an immersion solution during imaging that will match the refractive index of your sample. We will have a variety of buffers on hand, but these may not have a matching RI if your protocol is unusual.
Tissue clearing with organic solvents.Click here for a suggested protocol from LaVision, including advice on perfusion prior to tissue harvesting.
Free samples of a variety of rapid clearing solutions and matching RI gel made by SunJin Lab will be available during the workshop. Please see the SunJin Lab representative within TAC N223 who can also advise you on its use. The list of products offered can be found on the "Accessories" web page.