Hi Ted,

I really like Luigs and Neumann manipulators like SM-7 control system with Remote control SM7 / Keypad and  Mini 4 axes Unit. And Shifting table also from L&N for the microscope.

I think L&N is much stable than Sutter (currently I use a Sutter MPC-200 but I don't like it).

I hope it helps. Good luck

Mate

Hi Ted,

Here are some suggestions

MANIPULATORS: I agree with Mate, I've tried several brands, including Scientifica (which I find pretty good) and the L&N manipulators are better. I use SM7 as well.

MICROSCOPES: I found Olympus was so far the most satisfactory (compared to different Zeiss). I currently work with an Olympus BX51WI, which is equipped with DIC and with the 20x objective XLUMPLFLN20XW (NA = 1.0), and1x, 2x and 4x multiplying module mounted on top. With this combination the images are really neat and I could see cells deep in the brain slices as I never could with other systems. 

STAGE: (Post & Platform sample plate) and recording chamber are from Scientifica, non-motorized and very simple. It does the Job. I'm sure there's better around for more money. Other stages I used were all custom-built.

HEATING: I use an NPI system. I tried both heating the bath chamber and heating the inlet: the second solution is better as allowed a more precise temperature control, although it only works with small volumes of ACSF in the recording chamber (which is good for oxygenation and whashouts). The system is equipped with a thermometer that gives live feedback to the amplifyer. This way you can store the temperature fluctuations on your acquisition file. To me, this was very useful optimizing the system for recording spontaneous AP firing in brain slices (Cerebellum and substantia nigra) where firing rates are highly temperature-dependent. Over time, I found many unsuspected causes for drifts in temperature and prevented artifacts.

STIMULUS ISOLATORS: The cheapest and simplest isolator I found so far is Digitimer Ltd, NL800A. Batteries last for years but must be checked periodically: when they are running low, the output changes. I'm curretnyl using a WPI, SYS-A365R which I find better (but it costs three times as much).

I send you attached a list of some products and prices, based on recent quotations (prices are in Euro). Perhaps prices are different in the States but I hope it helps. Take care and good luck!

Bruno

P.S. Most of the companies agree to loan their equipment for a test-period (at least here in Austria).

P.P.S. To assemble the station, start simple, with as little equipment as possible, and make sure it's stable, noise-free and grounded without loops. Add the next components one by one. Large components may have parts to ground separately, such as the lower and  higher parts of my microscope. It is usually not possible to ground all the manipulator's columns without having ground loops. Careful checking with a multimeter is necessary. On the microscope, test the unpainted points such as screw holes, that can be used as grounding point as well.

Sutter instrument's manipulators 225/265/285 series and are quite stable and suitable. Also it's possible to use one control unit for up to 4 manipulators. Also Narishige's motorized mechanical manipulators are good, but not hydraulical. Eppendorf's micromanipulators are very unstable even for whole cell recordings.

Axon Axopatch amplifiers series 200 are good for voltage clamp, but may have big instrumental errors in current-clamp mode. But they are very simple and easy to use.

Hey Ted,

here are my preferences/suggestions based on working experience with these instruments.

Manipulators:  I have used Eppendorf (Patchman NP2), Sutter instrument and L&N, our current rigs have L& N (SM-5 and newer SM-7). Amongst these, L& N manipulators are the most stable and drift-free.

Microscope:We have (BX51Wl, Olympus) equipped with a 20x water-immersion objective (slice recording) and another rig is equipped with Axiovert 100 upright microscope (for single cell recording)

Recording chamber and platform:We have tried RC25 series with PM stages from Warner. These are quite nice if you intent to bath apply pharmacological agents.

Heating System:Thermoclamp-1 is a personal favourite, as it is affordable and works equally good as costly counterparts ( from warner instruments).

Hope it helps ! good luck for your set up.

Best 

Nand

Hi Ted, 

I used to work with Olympus BX51WI, it is very convinient and stable. As I remember some modifications have automatic objective changer and positioning.

Now I use Nikon Eclipse FN1 which is highly customizable and prety stable. It doesn't have separate metal parts so you can ground it easily. The only part which is sometimes inconvinient is a specific sliding objective changer. 

As for manipulators I would suggest L&M SM7 for its steadiness and handy control panel. I also use Sutter MP-225 for Cell Attached recordings and I can say that it is quite reliable too.

Bruno gave good advices in P.P.S. I would add that grounding tests are very important. One should check each metal thing near the setup, cause if something is not grounded properly it will become an aerial for 50-60Hz EMF. In my case once it was (intriguing pause) front sliding wall of Faraday cage.

I hope it helps. Good luck.

Tigran

Is this for whole-cell somatic patching in brain slices? Adding to what others have said above:

Olympus BX51WI body with tilting nosepiece (Olympus 4x/0.17 NA for targeting a particular area, and also for epi-illumination for imaging and ChR2 stimulation of UV uncaging; Olympus 60x/1.0 NA for approaching pipette to soma), fixed on air table. To visualize fluorescent neurons, etc. add the Olympus epi-illumination component with turret, various fluorescence cubes, and a suitable light source: Thorlabs sells multi-channel LEDs with adapters for the Olympus port, but I prefer to assemble these myself from Thorlabs parts - as this allows better adjustment of the light source position, to achieve true "inverse Koehler" illumination and thus even illumination at the sample.

Custom x/y-movable stage (cut a Thorlabs breadboard to shape/size, mount this on top of two posts with spherical bearings, and an x/y-translator on two more posts). Either manual x/y translation, or remote operation via high-current version of Sutter manipulator driver. With the latter, can also add a stepper motor for remote control of z-focus - very handy for precisely determining depth below slice surface, and to minimize mechanical disruption during patching.

Custom recording chamber. Inflow heating (Warner, TC324B) - make sure to insulate the inflow tubing and keep it very short. Sutter manipulators (MP-225, ROE-200, MPC-200), never had a drift problem with these (but only used for somatic recordings). Multiclamp 700B amplifier, National Instruments NI6259-USB DAQ.

All visualization via camera (no eye pieces), Retiga 2000R from QImaging. Instead of DIC, a cheaper (and very effective) method is oblique illumination with an IR LED (positioned below and to the side of the chamber) and a diffuser layer below the chamber. 

All of the above (plus laser scanning components) controlled by open-source Ephus software. Additional part #'s in Methods (see attached).

Article Ephus: Multipurpose Data Acquisition Software for Neuroscien...

Article Intrinsic Electrophysiology of Mouse Corticospinal Neurons: ...

Article Reciprocal Interareal Connections to Corticospinal Neurons i...

I want to thank everyone for their time and input. All of it was very helpful.

Peace,

Ted

Dear Ted

We have 3 rigs, all with Olympus scopes with DIC/IR as well as fluorescence; the latter is handy if you want to record from labeled cells.  The Olympus scopes have been excellent, as has the service.  You also need a fixed stage with a chamber (ours are from Siskiyou), an in-line bath heater and a good camera with controller and TV monitor.  We use the Multiclamp hardware/software, and like it.

We have manipulators from Siskiyou, which are digital and inexpensive (all relative here), and they are OK.  We also have a Sutter analog manipulator, which is awesome but very expensive.  

Good luck, and contact us if you have any questions.