I recommend using an air-impermeable oil like Paratone over the hassle of
capillaries. People often say that oil kills their crystals, but in my own
experience, that rarely happens. I think it is more likely due to
mishandling as working with oil does require some practice.
Once you get used to it
It¹s not just possible, but desirable! There¹s evidence that
conformational ensembles can be accessed in RT crystallography data, but
that these are frozen out in cryo data (aforementioned Fraser et al.,
Accessing protein conformational ensembles using room-temperature X-ray
crystallography. PNAS,
On 2/6/14, 10:32 AM, Thomas, Leonard M. wrote:
Finally as an observation over the years the art of mounting a crystal for a
room temperature or 4 C shot is a dying art. I am not firmly convince the
Mitogen mounts work really well, I have always had better luck with a
capillary. That just may
Room temperature data collection
Clearly, it is always possible to do non-cryogenic data collection simply by
not using a cryogenic cooling device and mounting crystals so that they do not
dehydrate or dry out.
I've been doing quite a lot of room temperature data collection lately becau
...@gmail.com]
Sent: Thursday, February 06, 2014 9:43 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Room temperature data collection
To answer Mark's question, if your crystals are capillary mounted then planes
are no problem. My protocol is to mount them, wrap them in cotton batting, put
them
To answer Mark's question, if your crystals are capillary mounted then
planes are no problem. My protocol is to mount them, wrap them in cotton
batting, put them in a 50 ml falcon tube, and pack that in bubble wrap. I
managed to get them through security in my carry-on with no issues. I
suggest usi
Dear Therese,
Yes, it is possible. If you want to collect data without cryo device on
an ESRF beam line (ID23 or ID29, all equipped with PILATUS now), you
need to remove 3 screws to remove the cryo device... We have as well 2
HC1 dehumidifiers which can be used on BM14 (which is equiped with a
I wonder whether flash-cooling from -10 degC would preserve those low mosaicity
values?
JPK
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of mesters
Sent: Thursday, February 06, 2014 8:40 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Room temperature data collection
Hello Theresa,
long time ago, we were having severe problems freezing crystals of
a human carboxypeptidase (GCPII). At room temperature and at 4
degrees celcius (capillary mounted), we could only collect 2 to 3
images at X13 in Hamburg and that was it (we h
Dear all,
Thermal motion reduction and lower radiation damage are reasons for
improved data associated
with low temperature data collection. The liquid to glass transition of
the solvent allows us to
have a better idea of the hydration shell around the protein. This is
something that room t
Clearly, it is always possible to do non-cryogenic data collection simply by
not using a cryogenic cooling device and mounting crystals so that they do not
dehydrate or dry out.
I've been doing quite a lot of room temperature data collection lately because
in the home lab we can SAD-phase lysoz
on a related matter, what are current experiences with taking crystals at room
temperature by plane?
(we are interested in trying the HC1 at a synchrotron but the closest are at
ESRF, which is quite a drive or train-ride from Madrid)
Mark J van Raaij
Lab 20B
Dpto de Estructura de Macromoleculas
Dear Enrico,
It is true that, on our beamline (FIP, at the ESRF), in situ (RT) is
mostly used for screening. But there is a fraction of cases where
freezing, and crystal handling, induces too much degradation. In these
cases, RT experiment is a real alternative.
In addition, data at room temp
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HI J,
do modern synchrotrons overcome the increase of thermal motion with
increasing temperature? Isn't this the main reason data are better
when collected at low temperature? Radiation damage is one reason for
sure, but not really the main reason. ;-
Dear Theresa,
We offer the possibility for data collection at room temperature on our
beamline, FIP-BM30A at the ESRF, for many years. It's really routine now
(see http://www.fip-bm30a.fr/).
Data are collected in situ, on the crystallization plate, thanks to the
robotized system we developed f
Dear Enrico,
"almost always it will be possible to achieve
better diffraction using cryogenic data collection."
I would say "almost always until now". Times change, instrumentation improves
and data collection techniques are becoming cunning. It's right time people
start
exploring the new poss
Dear Joern and other BBers,
While I fully agree that it is important to test a few images at room
temperature, to know the crystal's
potential, I think that almost always it will be possible to achieve
better diffraction using cryogenic
data collection.
Those rare cases, as the one you menti
Hi Theresa,
Diamond Light Source currently offers this capability at all of its
beamlines (all equipped with Pilatus detectors).
This can be performed either standard pins under controlled humidity
conditions, via the HC1 (Sanchez-Weatherby et al. *Acta Cryst.* (2009). D
*65*, 1237-1246 http://scri
Dear Theresa,
We recently collected a room temperature data set from one single crystal at
Petra III. The beam line was equipped with a Pilatus detector. Data were good
to 2.7 A. In contrast, at 100 K similar crystals diffracted very poorly. So, it
is perfectly possible to obtain useful room te
Dear crystallographers
Just out of curiosity, is it possible to collect datasets from crystals at room
temperature at synchrotron? Are fast detectors like Pilatus useful for this?
Thank you.
Theresa
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