Recent Posts

51

Surgery and Baseplating / Re: nothing but gray

« Last post by Daniel Aharoni on July 16, 2018, 05:43:29 PM »

Thanks for the schematics. This clarifies things a bit. So you actually just want to use 3 of those GRIN lenses stacked end to end with no gaps in between them.  This will effectively make a single GRIN lens with a 0.75 pitch which will work for imaging with a short working distance of ~50um to 250um.

52

Surgery and Baseplating / Re: nothing but gray

« Last post by hsiao on July 14, 2018, 09:27:01 AM »

Hi Daniel,

Thank you for your fast reply

I am currently using a setup like attached file Fig1A.
What you mean is I need to use a setup like Fig2A or 2B?

53

Surgery and Baseplating / Re: nothing but gray

« Last post by Daniel Aharoni on July 14, 2018, 06:33:22 AM »

Hi hsiao,
Can you clarify what you mean by stacking 2 objective lenses? Do you mean you put two of the #64-519 Edmund lenses end to end? If so this will create a ~0.5pitch GRIN lens which has very different optical properties than what the Miniscope is expecting as an objective lens. In other words, you have to keep the pitch of the objective lens to ~0.25 + 0.5*N pitch where N can be 0, 1, 2, ... We have imaged in rats by stacking 3 objective lenses together but using only 2 won't work.

 I am guessing the reason you saw clear signals outside the brain with a stack of 2 objective lenses is because you were imaging with a rather large working distance, a working distance much larger than what would work in the brain.

54

Surgery and Baseplating / nothing but gray

« Last post by hsiao on July 13, 2018, 06:28:56 AM »

Hi,

I was trying to do my first Gcamp experiment.

I planned to record dCA1 in rats. Since the grin lens (#64-519, Edmund optics) is not long enough, I stacked 2 lenses and screwed them in the bottom of Miniscope as an object lens. Then, I injected AAV into the dCA1 around the stratum radiatum and implanted 1 lens on the pyramidal layer (total 3 lenses, including 2 object and 1 relay lenses). After 3 weeks, I saw nothing. I even can’t see the blood vessels. I could only see black and gray under the maximal power and gain. I waited another week but still had no luck. You can check the screenshot below. This is a screenshot after 4 weeks of incubation.

 
I just perfused this rat, no obvious blood clot is noticed. I am still waiting for the histology and want to make sure my Gcamp express well but can’t wait to ask questions here for improving the protocol for my second rat.

 

I am not sure if my setup is correct (2 object lenses plus 1 relay lens) but I did test and see clear signals on a paper with green lines. I used some methods like this http://miniscope.org/index.php/Initial_Testing_of_Assembled_Miniscopes

 

Thank you!

55

DAQ Software/Firmware / Re: "Screen Shot" and "Record" Functions not Working Properly

« Last post by Daniel Aharoni on July 12, 2018, 05:13:05 PM »

Hi Chancey,
I'm not sure exactly what would be causing this. Make sure you are running the .exe file on a local hard drive and not over a network. Maybe try it out on another computer to make sure you can get it functioning properly somewhere.

56

DAQ Software/Firmware / "Screen Shot" and "Record" Functions not Working Properly

« Last post by chanceygarrett on July 12, 2018, 03:52:43 PM »

Hello,

I have the miniscope system up and running smoothly with no issues except that I cannot save data files from the software. Images come up nicely once I connect this miniscope to the software. When I hit the "record" or "screen shot" buttons in the DAQ software I received from github.com, the message board at the bottom right of the screen says it is saved, but when I look in the folder with the executable file in it, there is nothing. I've searched for data files through the entire computer, and I have tried uninstalling and reinstalling the software multiple times with no change. What do you think is occurring, what can I change to fix this?

Thanks!
Chancey

57

Optics / Re: Calibration

« Last post by Daniel Aharoni on July 10, 2018, 08:59:59 PM »

Hi Ryan,
The relevant GRIN lens information you need can be found here http://miniscope.org/index.php/GRIN_Lens_Information and here http://miniscope.org/index.php/Imaging_With_Thin_GRIN_Lenses.

58

Optics / Re: Calibration

« Last post by ryancameron on July 10, 2018, 05:53:04 PM »

Okay, that's probably our problem. What specs are the objective GRIN lenses usually? And will that length change depending on the size of our implanted (in the animal) lens?

Thank you!
Ryan Cameron

59

Optics / Re: Calibration

« Last post by Daniel Aharoni on July 10, 2018, 07:11:04 AM »

Hi Ryan,
Just to clarify, are you trying to imaging with only these thin (1mm diameter or less) GRIN lenses or are you imaging in conjunction with a larger (1.8 or 2mm diameter) objective GRIN lens. You need the objective GRIN lens in place to see anything.

60

Electronics / Re: Using a sony sensor with the miniscope DAQ?

« Last post by Daniel Aharoni on July 10, 2018, 07:09:39 AM »

Hi,
I haven't had the chance to look over those sensors' data sheets but you will want to find imaging sensors that output an 8 to 12bit parallel bus with clock, line valid, and frame valid control signals all running at under 100MHz. Many large format sensors output over multiple LVDS lines which the rest of the Miniscope electronics cannot handle currently.