thanks! ... Fwd: Re: has anyone put these wickedlasers (or equivalent price/power) on confocal, MSIM, single molecule localization etc scope?
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George McNamara |
thanks! ... Fwd: Re: has anyone put these wickedlasers (or equivalent price/power) on confocal, MSIM, single molecule localization etc scope?
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi David, Thanks! I checked out Viasho which led me to two USA distributors. Here's two lasers and price that caught my eye (and hopefully will never be pointed at mine or anyone else's): http://www.lasernrg.com/diode-and-dpss-laser-modules.html *20W/445nm laser module <http://www.synchrovision.co.uk/product/179-20w-445nm-laser-module-kvant>* EUR 6,356 *6.8W/637nm laser module <http://www.synchrovision.co.uk/product/175-6-8w-637nm-laser-module-kvant> *?EUR 10,625 (a bit weird that a USA distibutor web site would post prices in Euro's ... but the links are to a UK company). and, can apparently buy online: http://www.synchrovision.co.uk/product/175-6-8w-637nm-laser-module-kvant 6.8 Watts, even if 90% power loss getting to each objective lens, would imply 680 mW for one microscope (CW STED depletion and/or CW 2-photon excitation of DAPI), or 68 mW for 10 scopes (and $1065 per laser, ignoring splitters, intensity control - and safety goggles). I am interested in setting up an instrument to ablate single cells. 20W/445nm should do it, though I might be concerned with also ablating the optics and ceiling (latter assuming an inverted stand). Another reason I am thinking about relatively low cost lasers is the potential to provide light to new nanoscope(s), such as ... York ... Shroff 2012 "MSIM" paper in Nature Methods: For exciting fluorescence, two lasers were used: a 150-mW, 561-nm laser (561, Coherent, Sapphire 561-150 CW CDRH) and a 200-mW, 488-nm laser (488, Coherent, Sapphire 488-200 CDRH). Mechanical shutters (Thorlabs, SH05 and SC10) placed after each laser were used to control illumination. Beams were combined with a dichroic mirror (DC, Chroma, 525dcxru) and expanded 6.7× with a beam expander constructed from two achromatic lenses (Edmund, f = 30 mm, NT49-352-INK and Thorlabs, f = 200 mm, AC254-200-A-ML). Expanded beams were directed onto a DMD (Digital Light Innovations, D4100 DLP 0.55" XGA) 24 degrees off normal, so that in the 'on' position the micromirrors tilted the output beam normal to the DMD face. The central order of the resulting pattern was demagnified 1.5× with a beam de-expander (Thorlabs, f = 75 mm, AC254-075-A-ML and f = 50 mm, AC254-050-A-ML), aligned in a 4f configuration such that the DMD face was reimaged at the back focal plane of a 180 mm tube lens internal to the microscope (Olympus, IX81). Probably would not need multi-Watts, so something like: *200mW/405nm laser module <http://www.synchrovision.co.uk/product/87-200mw-405nm-laser-module-kvant>*EUR 770 (100 mW is EUR462, but why bother) *200mW/445nm laser module <http://www.synchrovision.co.uk/product/78-200mw-445nm-laser-module-kvant>*EUR 540 *300mW/532nm laser module <http://www.synchrovision.co.uk/product/76-300mw-532nm-laser-module-kvant>*EUR 739 *170mW/637nm laser module <http://www.synchrovision.co.uk/product/71-170mw-637nm-laser-module-kvant>*EUR 406 or *150mW/642 (640) nm laser module <http://www.synchrovision.co.uk/product/61-150mw-642-640-nm-laser-module-kvant>*EUR 394 Or, Xiaowei Zhuang's latest - eight standard organelle dyes are STORM-able, http://www.pnas.org/content/early/2012/08/06/1201882109.abstract (open access article) Sincerely, George -------- Original Message -------- Subject: Re: has anyone put these wickedlasers (or equivalent price/power) on confocal, MSIM, single molecule localization etc scope? Date: Sun, 19 Aug 2012 16:52:27 -0700 (PDT) From: David Baddeley <[hidden email]> Reply-To: David Baddeley <[hidden email]> To: [hidden email] <[hidden email]> I can't find any details about the actual design of the laser modules the website, but I looked into getting a very similar 1W blue laser pointer that was on offer at deal-extreme. It turned out that the module was originally designed for laser projection/lighting applications and was a number of separate lower wattage modules fabricated side by side on the same die (like some high power LEDs). This type of laser is unsuitable/less suitable for microscopy applications due to the extended source area and corresponding inability to be focussed down to a small spot. To switch / control intensity your best bet would probably be an external shutter and/or filter wheel. We use viasho lasers which are positioned, both in price/W and capabilities/quality, somewhere between these and classical 'scientific' grade lasers. The viasho modules have either TTL or analog control via bnc, but switching the lasers using these inputs results in instability and mode hopping as they warm up and cool down (I suspect the inputs are designed for more rapid modulation on timescales less than the thermal relaxation of the laser die), making shutters and a filter wheel the most practical method of control. As to the glasses, they sell them separately as OD2 glasses without specifying which of their lasers they should be used with, so I suspect they've just specced them to be OD2 or better at all wavelengths of laser they sell. This actually seems like a fairly responsible/ pragmatic approach for people selling a product like this. cheers, David ------------------------------------------------------------------------ *From:* George McNamara <[hidden email]> *To:* [hidden email] *Sent:* Sunday, 19 August 2012 1:23 AM *Subject:* has anyone put these wickedlasers (or equivalent price/power) on confocal, MSIM, single molecule localization etc scope? ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear listserv, has anyone put these wickedlasers (or equivalent price/power) on confocal, MSIM, single molecule localization etc scope (and figured out how to control on/off, intensity, from USB or other port)? http://www.wickedlasers.com/krypton?utm_source=Wicked+Lasers+New&utm_campaign=bd89072d39-S3_August_20128_17_2012&utm_medium=email <http://www.wickedlasers.com/krypton?utm_source=Wicked+Lasers+New&utm_campaign=bd89072d39-S3_August_20128_17_2012&utm_medium=email> 532 nm, 750 mW, $999.95, Spyder Krypton 445 nm, 1250 mW, $399.95, Spyder Arctic If you have, web page or other instructions, on how would be great. George p.s. if someone can explain to me (without cheating by asking the company) the choice of glasses in the home page video, http://www.wickedlasers.com/index.php , that would be great. If you make it through the first video, click on the Arctic Popcorn video link. And of course, "do not look at laser with remaining eye". |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Listers, I have a Nikon A1 with the spectral detector. Last week, yes on Friday at 4pm, I had some users trying to capture the spectral output of a weak silicon chip with some sort of metals on the surface with current running through it. I could view the light (orange to red) at both 10x and 20x through the oculars, but could not figure out how to configure the A1 to detect the photons. Of course there are plenty of set ups which are far better designed for this application, but that's not my question. My question is simply how best to detect several wavelength (between around 600 and 750nm) of light being emitted on the A1 or if it is possible. I was able to get a very weak and pixelated image with one set of setting, and a huge blob of photos with just the standard "Cy-5" via the normal PMT's. Ideas? Ultimately, the PI would like to view the light emitting areas and characterize the wavelengths coming from those areas. Thanks. Christian |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi Christian, Am I correct in assuming this was without any laser illumination? That is, "electroluminescence" (I forget the official phrase). If this is correct, then you are using the scanning mirrors to create the image. Since you are not using a point illumination source (the focused lasers), you will probably be better off opening the pinhole to collect more light. However, on the spectral confocal systems I am familiar with (Zeiss LSM710 and Leica SP5 and SP1) opening the pinhole compromises the spectral resolution. To get a sense for how this behaves with more standard confocal specimens, a good test specimen is a bright DAPI labeled nucleus (nuclei) excited using the microscope arc lamp and DAPI filter set. This test might help you compare behavior of the spectral vs standard PMTs in "not a laser illumination" specimen. George On 8/19/2012 9:10 PM, Christian wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Listers, > > I have a Nikon A1 with the spectral detector. Last week, yes on Friday at 4pm, I had some users trying to capture the spectral output of a weak silicon chip with some sort of metals on the surface with current running through it. I could view the light (orange to red) at both 10x and 20x through the oculars, but could not figure out how to configure the A1 to detect the photons. Of course there are plenty of set ups which are far better designed for this application, but that's not my question. My question is simply how best to detect several wavelength (between around 600 and 750nm) of light being emitted on the A1 or if it is possible. I was able to get a very weak and pixelated image with one set of setting, and a huge blob of photos with just the standard "Cy-5" via the normal PMT's. > > Ideas? Ultimately, the PI would like to view the light emitting areas and characterize the wavelengths coming from those areas. > > Thanks. > > Christian > > |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** On 8/20/12 4:45 AM, George McNamara wrote: > > If this is correct, then you are using the scanning mirrors to create > the image. Since you are not using a point illumination source (the > focused lasers), you will probably be better off opening the pinhole to > collect more light. However, on the spectral confocal systems I am > familiar with (Zeiss LSM710 and Leica SP5 and SP1) opening the pinhole > compromises the spectral resolution. > In the Nikon C1 and A1 spectral, the spectral unit is on the other side of a fiber, while the pinhole is in the scanhead. Therefore, spectral resolution should be independent of pinhole size. --aryeh -- Aryeh Weiss Faculty of Engineering Bar Ilan University Ramat Gan 52900 Israel Ph: 972-3-5317638 FAX: 972-3-7384051 |
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In reply to this post by George McNamara
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** I'm not terribly familiar with the A1 but in general I'd suggest the following approach. Acquire a confocal (reflection or fluorescence) of the device and move the sample to position one of the regions of interest at the centre of the frame. Park the beam at this spot and turn off the laser, open the pinhole, turn on the current to the device and acquire a spectrum for as long as it takes. (Could be hours). Repeat for the other light-emitting areas. In other words, separate the collection of the image and the spectrum. Having said that, you can get a pocket spectrometer from Ocean Optics, with a microscope adapter, for a very low price. This will do the job much more simply and effectively. And don't forget that you'll need metallurgical objectives if you want to get a decent image. Guy -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of George McNamara Sent: Monday, 20 August 2012 11:45 AM To: [hidden email] Subject: Re: odd use of an A1 ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi Christian, Am I correct in assuming this was without any laser illumination? That is, "electroluminescence" (I forget the official phrase). If this is correct, then you are using the scanning mirrors to create the image. Since you are not using a point illumination source (the focused lasers), you will probably be better off opening the pinhole to collect more light. However, on the spectral confocal systems I am familiar with (Zeiss LSM710 and Leica SP5 and SP1) opening the pinhole compromises the spectral resolution. To get a sense for how this behaves with more standard confocal specimens, a good test specimen is a bright DAPI labeled nucleus (nuclei) excited using the microscope arc lamp and DAPI filter set. This test might help you compare behavior of the spectral vs standard PMTs in "not a laser illumination" specimen. George On 8/19/2012 9:10 PM, Christian wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Listers, > > I have a Nikon A1 with the spectral detector. Last week, yes on Friday at 4pm, I had some users trying to capture the spectral output of a weak silicon chip with some sort of metals on the surface with current running through it. I could view the light (orange to red) at both 10x and 20x through the oculars, but could not figure out how to configure the A1 to detect the photons. Of course there are plenty of set ups which are far better designed for this application, but that's not my question. My question is simply how best to detect several wavelength (between around 600 and 750nm) of light being emitted on the A1 or if it is possible. I was able to get a very weak and pixelated image with one set of setting, and a huge blob of photos with just the standard "Cy-5" via the normal PMT's. > > Ideas? Ultimately, the PI would like to view the light emitting areas and characterize the wavelengths coming from those areas. > > Thanks. > > Christian > > |
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In reply to this post by Christian-103
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** I do not have A1, but this may work: I would switch to the lowest spectral resolution (10 nm per channel), open the pinhole, and on the beamsplitter select a dichroic mirror that transmits in the orange/red (e.g., the standard 405/488 or 457/514, or even better if the 8- position turret has an empty slot without any filter, use that). of course turn the lasers off. the 20x objective may be a better choice than the 10x (higher NA, collects more photons). Do a slow scan, if necessary use averaging. if you do raster scan (instead of just a single spot scanning), use pixel spacing comparable to resolution, or bigger (i.e., do not try to do Nyquist sampling). Stan Vitha Microscopy and Imaging Center Texas A&M University On Tue, 21 Aug 2012 07:21:09 +0000, Guy Cox <[hidden email]> wrote: >***** >To join, leave or search the confocal microscopy listserv, go to: >http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >***** > >I'm not terribly familiar with the A1 but in general I'd suggest the following approach. Acquire a confocal (reflection or fluorescence) of the device and move the sample to position one of the regions of interest at the centre of the frame. Park the beam at this spot and turn off the laser, open the pinhole, turn on the current to the device and acquire a spectrum for as long as it takes. (Could be hours). Repeat for the other light-emitting areas. In other words, separate the collection of the image and the spectrum. > >Having said that, you can get a pocket spectrometer from Ocean Optics, with a microscope adapter, for a very low price. This will do the job much more simply and effectively. > >And don't forget that you'll need metallurgical objectives if you want to get a decent image. > > Guy > >-----Original Message----- >From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of George McNamara >Sent: Monday, 20 August 2012 11:45 AM >To: [hidden email] >Subject: Re: odd use of an A1 > >***** >To join, leave or search the confocal microscopy listserv, go to: >http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >***** > >Hi Christian, > >Am I correct in assuming this was without any laser illumination? That > >If this is correct, then you are using the scanning mirrors to create the image. Since you are not using a point illumination source (the focused lasers), you will probably be better off opening the pinhole to collect more light. However, on the spectral confocal systems I am familiar with (Zeiss LSM710 and Leica SP5 and SP1) opening the pinhole compromises the spectral resolution. > >To get a sense for how this behaves with more standard confocal specimens, a good test specimen is a bright DAPI labeled nucleus >(nuclei) excited using the microscope arc lamp and DAPI filter set. This test might help you compare behavior of the spectral vs standard PMTs in "not a laser illumination" specimen. > >George > >On 8/19/2012 9:10 PM, Christian wrote: >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> Listers, >> >> I have a Nikon A1 with the spectral detector. Last week, yes on Friday at chip with some sort of metals on the surface with current running through it. I could view the light (orange to red) at both 10x and 20x through the oculars, but could not figure out how to configure the A1 to detect the photons. Of course there are plenty of set ups which are far better designed for this application, but that's not my question. My question is simply how best to detect several wavelength (between around 600 and 750nm) of light being emitted on the A1 or if it is possible. I was able to get a very weak and pixelated image with one set of setting, and a huge blob of photos with just the standard "Cy-5" via the normal PMT's. >> >> Ideas? Ultimately, the PI would like to view the light emitting areas and characterize the wavelengths coming from those areas. >> >> Thanks. >> >> Christian >> >> |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Thank you all for the input. I understand this is not really the best use for the instrument, but we often use what we have for what is needed. --- On Wed, 8/22/12, Stanislav Vitha <[hidden email]> wrote: From: Stanislav Vitha <[hidden email]> Subject: Re: odd use of an A1 To: [hidden email] Date: Wednesday, August 22, 2012, 10:07 AM ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** I do not have A1, but this may work: I would switch to the lowest spectral resolution (10 nm per channel), open the pinhole, and on the beamsplitter select a dichroic mirror that transmits in the orange/red (e.g., the standard 405/488 or 457/514, or even better if the 8- position turret has an empty slot without any filter, use that). of course turn the lasers off. the 20x objective may be a better choice than the 10x (higher NA, collects more photons). Do a slow scan, if necessary use averaging. if you do raster scan (instead of just a single spot scanning), use pixel spacing comparable to resolution, or bigger (i.e., do not try to do Nyquist sampling). Stan Vitha Microscopy and Imaging Center Texas A&M University On Tue, 21 Aug 2012 07:21:09 +0000, Guy Cox <[hidden email]> wrote: >***** >To join, leave or search the confocal microscopy listserv, go to: >http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >***** > >I'm not terribly familiar with the A1 but in general I'd suggest the following approach. Acquire a confocal (reflection or fluorescence) of the device and move the sample to position one of the regions of interest at the centre of the frame. Park the beam at this spot and turn off the laser, open the pinhole, turn on the current to the device and acquire a spectrum for as long as it takes. (Could be hours). Repeat for the other light-emitting areas. In other words, separate the collection of the image and the spectrum. > >Having said that, you can get a pocket spectrometer from Ocean Optics, with a microscope adapter, for a very low price. This will do the job much more simply and effectively. > >And don't forget that you'll need metallurgical objectives if you want to get a decent image. > > Guy > >-----Original Message----- >From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of George McNamara >Sent: Monday, 20 August 2012 11:45 AM >To: [hidden email] >Subject: Re: odd use of an A1 > >***** >To join, leave or search the confocal microscopy listserv, go to: >http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >***** > >Hi Christian, > >Am I correct in assuming this was without any laser illumination? That > >If this is correct, then you are using the scanning mirrors to create the image. Since you are not using a point illumination source (the focused lasers), you will probably be better off opening the pinhole to collect more light. However, on the spectral confocal systems I am familiar with (Zeiss LSM710 and Leica SP5 and SP1) opening the pinhole compromises the spectral resolution. > >To get a sense for how this behaves with more standard confocal specimens, a good test specimen is a bright DAPI labeled nucleus >(nuclei) excited using the microscope arc lamp and DAPI filter set. This test might help you compare behavior of the spectral vs standard PMTs in "not a laser illumination" specimen. > >George > >On 8/19/2012 9:10 PM, Christian wrote: >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> Listers, >> >> I have a Nikon A1 with the spectral detector. Last week, yes on Friday at chip with some sort of metals on the surface with current running through it. I could view the light (orange to red) at both 10x and 20x through the oculars, but could not figure out how to configure the A1 to detect the photons. Of course there are plenty of set ups which are far better designed for this application, but that's not my question. My question is simply how best to detect several wavelength (between around 600 and 750nm) of light being emitted on the A1 or if it is possible. I was able to get a very weak and pixelated image with one set of setting, and a huge blob of photos with just the standard "Cy-5" via the normal PMT's. >> >> Ideas? Ultimately, the PI would like to view the light emitting areas and characterize the wavelengths coming from those areas. >> >> Thanks. >> >> Christian >> >> |
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