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Hello every one,
We are trying to use the deconvolutionJ plug-in to deconvolve an epifluorescent image stack, collected using the Zeiss 100X NA 1.4 NeoFluar objective and a Hamamatsu ORCA camera. Now we are wondering how to make the PSF file for the objective, which is esssential for the deconvolution. We will be grateful if someone could help us with a setp-by-step instruction, or, it would be even better if someone could supply the file. many thanks, krishanu. Krishanu Ray Department of Biological Sciences Tata Institute of Fundamental Research Homi Bhabha Road, Colaba, Mumbai 400 005. India. Ph: +91-22-22804545 ext 2730 Fax: +91-22-22804610/11 |
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Hello,
what do you need is to acquire a set of optical sections of microspheres fluo sample of 0.1 micron in diameter and to collect data at 0.1 micron or less step along z-axis. I suggest to start 10 microns below and to stop 10 microns above the focal plane using 512x512 and to follow the method described in Agard&Sedat paper: Biophysical Journal, Vol 57, 325-333 Determination of three-dimensional imaging properties of a light microscope system. Partial confocal behavior in epifluorescence microscopy Y Hiraoka, JW Sedat and DA Agard Giancarlo At 19.39 15/12/2005 +0530, you wrote: >Hello every one, > >We are trying to use the deconvolutionJ plug-in to deconvolve an >epifluorescent image stack, collected using the Zeiss 100X NA 1.4 >NeoFluar objective and a Hamamatsu ORCA camera. > >Now we are wondering how to make the PSF file for the objective, >which is esssential for the deconvolution. > >We will be grateful if someone could help us with a setp-by-step >instruction, or, it would be even better if someone could supply the file. > >many thanks, > >krishanu. > >Krishanu Ray >Department of Biological Sciences >Tata Institute of Fundamental Research >Homi Bhabha Road, Colaba, Mumbai 400 005. >India. >Ph: +91-22-22804545 ext 2730 >Fax: +91-22-22804610/11 ... [show rest of quote] Giancarlo Mascetti, Ph.D. X-Istituto di Calcolo Scientifico c/o Paramed Health Services Ltd. c.so F. M. Perrone, 73r - 16152 Genova, Italy tel. 010-6489265 - mob. 335-7787927 - fax. 010-7404530 E-mail: [hidden email] "Le informazioni contenute in questo messaggio di posta elettronica, unitamente ai suoi eventuali allegati, sono di natura riservata e per l'uso esclusivo del(i) destinatario(i) e solo per lo scopo riportato nel messaggio stesso. Se Lei non e' tra i destinatari, si rammenta che qualsiasi divulgazione, distribuzione, modifica o copia che Lei possa farne e' proibita. Pertanto se il presente messaggio e' ricevuto per errore, si prega di distruggerlo e di informarci immediatamente via e-mail. The present e-mail and any attachment files, contain company's confidential information and it is intended for the person to whom is addressed and for the purpose therein set forth, only. If you are not the intended recipient (or if you have received this e-mail in error) please notify the sender immediately and destroy this e-mail. Any distribution, dissemination, disclosure, copying or other use of this e-mail by person or entities other than the intended recipient is strictly forbidden." |
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