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
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
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 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
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 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
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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 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
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
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
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
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
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
He
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
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
If one is mounting in glass capillaries, we used to put them in a box of blue
tips (1000 ul pipetteman tips). Just put a small rolled up wad of kimwipe down
in the bottom, so that the capillary does not jam itself down in the narrowing
of the tip point. Every capillary gets its own tip and one
To add my 2 cents. Recently we have screened a lot of really poor looking
crystals. We had tried originally at cryo temps and really were quite unsure
as to weather they were protein, just poorly diffracting crystals, etc.. We
then mounted some at room temperature and were able to observed rea
Additionally, once the crystals are ready for the trip, remember to place a
heat sink in with the samples. Planes and tarmacs are seldom at room
temperature.
Kris
Kris F. Tesh, Ph. D.
Department of Biology and Biochemistry
University of Houston
From: Bernard
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
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,
All,
I was asked by the meeting organizers to let the CCP4 community know
about the Residential School on Medicinal Chemistry and Biology in Drug
Discovery.See below for information about the course and contact
information.
Sincerely,
Corey
--
Dear ACA attendees,
We would like to invite you to submit an abstract to our session:
3.2.5. Chemistry and Biology with Novel Scattering Techniques
scheduled from 1:30PM to 5:00PM on Tues., May 27, 2014 at the American
Crystallography Association meeting in Albuquerque. Our session is aimed
We recently published a method for estimating F(000) by simply equating
it to Fcalc(000), the sum of all the electrons in the refined model
(including those in the bulk solvent):
http://dx.doi.org/10.1073/pnas.1302823110
The results of this procedure compared favorably with the two
experimen
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
Post-doctoral Research Associate positions are available for the
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Hello,
Does anybody know what would be a good cryoprotectant for the following
condition:
800 mM Sodium phosphate monobasic/1200 mM Potassium phosphate dibasic 100
mM Sodium acetate/Aceticacid pH4.5
Thanks
Deepak
Lots of choices. I usually try the crystallization solution + 30% glucose
first. Glycerol or ethylene glycol are other possibilities here. Another
possibility is 2.5-3.0 M sodium malonate at a similar pH.
Roger Rowlett
On Feb 6, 2014 11:40 PM, "Deepak Thankappan Nair"
wrote:
> Hello,
> Does anyb
You might get some clues from
Acta Cryst. (2008). D64, 287-301[ doi:10.1107/S0907444907067613 ]
Glycerol concentrations required for the successful vitrification of cocktail
conditions in a high-throughput crystallization screen
R. Kempkes, et al.
and
J. Appl. Cryst. (2002). 35, 538-545
Try L-proline. It works well with high ionic strength conditions:
http://www.ncbi.nlm.nih.gov/pubmed/22868767
Sent from Jack's iPad
On Feb 6, 2014, at 10:40 PM, "Deepak Thankappan Nair"
mailto:deepaktn...@gmail.com>> wrote:
Hello,
Does anybody know what would be a good cryoprotectant for the
Dear all,
at the 23rd IUCr congress (http://www.iucr2014.org/), which will take
place in August this year in lovely Montreal, John Rose and I are
organizing the microsymposium MS-40 titled "S-SAD and other
applications of soft X-rays in MX".
If you happen to have an exciting story, which you wou
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