概要

國立陽明大學推廣神經科學的研究以及教育長達30年,本校的神經科學研究所成立於1980年,為台灣首個致力推廣神經科學領域的研究單位,而緊接腦科學研究所亦於2005年成立,目標為建立集基礎研究和臨床領域相輔相佐的研究單位。同時,台北榮民總醫院亦建立神經內科與神經外科研究所,致力實踐臨床神經科學和醫療的研究。而我置身於這蘊含著豐富的神經科學教育資源及臨床研究的環境下學習研究。指導教授的辦公室位於圖書資訊暨研究大樓,而實驗室的總面積為173平方米,其中包括了一個用於生物化學或分子生物的實驗室和一個培養房,另外還有三個用作電生理的房間,而主要的實驗器材如下:

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                                                                             

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

實驗室資源

(1) Full access to central facilities including the P2 lab (lentivirus and rAAV generation), genomics/proteomics core, imaging core facilities, animal behavioral core laboratory.

(2) All aliquots of live virus and MTA information: 
Virus in Lien Lab: [Link] 

(3) Optogenetic setups: We have two LED sources (one LED4D162 consisting of 420/470/505/590 nm, controlled by DC4104 driver, Thorlabs, NJ, USA; and one Doric 470 nm LED, controlled by Doric LED Driver-1000, Doric lenses Inc., Canada), and one laser light source (DPSS blue 473nm laser, OEM Laser Systems, USA) for activation of light-sensitive channels.

(4) Six brain slice patch-clamp recording setups (upright IR-DIC or IR-DGC microscopes; four Olympus BX51WI, Olympus, Tokyo, Japan; one Leica  DM6000 CFS, Leica, Wetzlar, Germany; one Zeiss Axioskop 2 F5, Carl Zeiss, Göttingen, Germany); each setup consists of at least two motorized micromanipulators (twelve Mini-25 Units and two Junior Units, controlled by five SM5, two SM6 and one SM7 Control boxes; Luigs & Neumann, Ratingen, Germany) or manual manipulators (LBM-7; Scientifica, UK), a CCD camera (two Hamamatsu C7500-50, one Hamamatsu C3077-79, one Hamamatsu C2400-79H, Hamamatsu Photonics, Japan; and one IR-1000, Dage-MTI, USA) and a LCD or a CRT monitor.

(5) An extracellular field potential recording rig; the setup consists of an interface recording chamber (BSC2 Brain Slice Heated Chamber, Scientific Systems Design Inc., Digitimer, UK), one proportional temperature controller (PTC03, Scientific Systems Design Inc., Digitimer, UK), one extracellular amplifier (EXT-02F, NPI Electronic Instruments, Tamm, Germany), three manual manipulators (LBM-7; Scientifica, UK), one motorized micromanipulator (MicroStar; Scientifica, UK)

(6) Five Multiclamp 700B amplifiers, six pClamp software and four Axon patch 200B amplifiers (Molecular Devices, CA, USA)

(7) Seven digitizers (six 1440A Digidata; one 1332A Digidata; Molecular Devices, CA, USA)

(8) Five oscilloscopes (two Tektronix TDS 1002, USA; one Tektronix, GDS 620, USA; one Iwatsu SS-7810 1002, Iwatsu Test Instrument Corporation, Tokyo, Japan; and one GW Instek GOS-620, Good Will Instrument, Taiwan)

(9) Eight stimulation isolators (ISO-flex, A.M.P.I, Israel), one Master 8 programmed stimulator (A.M.P.I, Israel), and one isolated pulse stimulator (model 2100, A-M Systems, USA)

(10) Three heating systems (two Temperatur controller TC05, Luigs & Neumann, Ratingen, Germany; and one single channel heater controller, TC-324B equipped with a SH-27B solution on-line heater, Warner Instrument Corp., USA)

(11) A Metamorph software (v.7.5.2.0; Molecular Devices, CA, USA) and a Neurolucida  reconstruction software (10.40 64-bit; MicroBrightField, Williston, VT, USA)

(12) Six stereoscopic microscopes (three Olympus SZ61, Olympus, Tokyo, Japan; one Leica EZ4D, Leica Microsystems, Germany; one Labophot 2, Nikon, Tokyo, Japan; one Olympus BH2 microscope, Olympus, Tokyo, Japan )

(13) One confocal/two-photon laser excitation microscope (Leica SP5 module, Leica Microsystems, Germany)
Objective lens/物鏡 :
Dry: N PLAN 5x/0,12
Water immersion: HC APO L 20x/0,50 W U-V-I
HC APO L 63x/0,90 W U-V-I
Laser: Argon (458 nm, 476 nm, 488 nm, 514 nm)
HeNe 543 nm
HeNe 633 nm

(14) Three tissue microslicers (DTK-1000, Dosaka, Kyoto, Japan)

(15) One osmometer (Osmomate 030, Gonotec, Berlin, Germany)

(16) One pH meter(Seven Easy S20, Metter Toledo, Greifensee, Switzerland)

(17) One Millipore pure water system(Direct-Q 3UV water purification system, Millipore, Molsheim, France)

(18) Two sets of immunoblot devices (Mini Trans-blot cell, Bio-Rad, Hercules, CA, USA )

(19) One PCR thermal cycler (Labcycler; SensoQuest GmbH, Göttingen, Germany)

(20) Three glass pipette pullers (one Sutter P-97, Sutter Instruments, Novato, CA, USA; two PC-10, Narishige, Tokyo, Japan)

(21) Four PicoSpritzer III (Parker Instrumentation, Pine Brook, NJ, USA), and one  pneumatic picopump (PV830, World Precision Instruments, Sarasota, FL, USA)

(22) Three microforges (two MF-830, Narishige, Tokyo, Japan; one MF-200, World Precision Instruments, Sarasota, FL, USA )

(23) For ultra-fast solution exchange: We have two piezoelectric actuators (one Burleigh LSS-3000, EXFO, Ontario, Canada; and one MXPZT-300, Siskiyou, USA)

(24) One microperfusion system (SmartSquirt Micro-Perfusion System, AutoMate Scientific, Berkeley, CA )

(25) Nine peristaltic pumps (three Perista pump SJ-1211, ATTO, Tokyo, Japan;  two ISM 834C, Ismatec, SA, Switzerland; two Miniplus Evolution, Gilson, USA; one BT100-2J, Longer Pump, China, one Alitea SC, Watson–Marlow, Alitea AB, Sweden)

(26) Two infusion pumps (KD-230, KD Scientific, Boston, MA, USA; Fusion 100, Chemyx, Stafford, TX, USA)

(27) One culture hood (1300 Series A2 Class II, Thermo Scientific, Asheville, NC, USA)

(28) One CO2 incubator (Forma 310 Series Direct Heat CO2 Incubator, Thermo Scientific, Asheville, NC, USA)

(29) One chemical hood

(30) Two ovens/two ultrasound sonicator

(31) Three freezers (-70 ˚C,-40 ˚C, and -20 ˚C) and one fridge (4˚C)

(32) Ten Vacuum pumps (Tecnomara AG, Wallisellen, Switzerland)

(33) One centrifuge (Microcentrifuge 5415R, Eppendorf, Hamburg, Germany)

(34) One Research High-Class Stereo Microscope System (SZX16, Olympus, Tokyo, Japan) with Reflected  Flurescence System.
Stereo microscopes: Olympus SZX16
Digital Microscopy Camera: Olympus DB80
Filter:
For UV excitation: Ex330-385/Em 420- (SZX2-FUV)
For GFP: Ex460-490/Em510- (SZX2-FGFP)
For RFP: Ex530-550/Em575- (SZX2-FRFP1)
Power supply: Olympus U-RFL-T

(35) Ca2+ imaging setup system in Lien’s lab

        Light source: Polychrome V (150 W Xenon lamp, 320-694 nm, bandwidth ~15 nm, TILL Photonics GmbH, Germany)

        Optical fiber: Quartz light guide (NA 0.22) with an epi-uorescence condenser (TILL Photonics GmbH, Germany)

        Coupler to microscope (light source): BX-RFA (Olympus, Japan)

        Neutral density lter: ND-25 (Olympus, Japan)

 

        Filter sets:

       

Mirror unit

Dichroic mirror

Excitation filter

Barrier filter

U-MWU2

DM400

BP330-385

BA420

U-MWIB3 (for OGB-1)

DM505

BP460-495

BA510IF

U-MWIG3

DM570

BP530-550

BA575IF

Fura-2 (customized)

400DCLP

empty

E420LPv2

       *Filter sets for Fura-2 were purchased from Chroma Technology Corp., USA; others from Olympus, Japan.

        Camera: EM-CCD camera (QuantEM 512SC, Photometrics, USA)

        Coupler to microscope (camera): Dual Photo Port (U-DPT, Olympus, Japan), C-mount (U-CMAD3, Olympus, Japan)

        Controlling software: MetaFluor (Molecular Devices, USA)

Interface: Breakout box (for light source), RJ-45 (attached to Polychrome V), RS-232/BNC (attached to PC) (TILL Photonics GmbH, Germany), IEEE-1394a (for EM-CCD), (Turbo-1394™ interface, Photometrics, USA), LTP port controlled box (custom made, for digitizer), BNC (attached to digitizer), LTP port (attached to PC)

PC platform:CPU, 2.4 GHz, Intel® Core™2 Quad (Intel, USA), RAM 2 GB, Graphics card Geforce 8500 GT (NVIDIA, USA)

Operating System, Windows XP sp3 (Microsoft, USA), Mother board with RS-232, IEEE-1394a and LTP sockets

(36)  Voltage imaging equipment

Upright microscope: BX51W1; Olympus

Light source: 532 nm diode-pumped solid-state laser; 500 mW; OEM Laser System

CCD camera: NeuroCCD-SMQ; RedShirtImaging

Shutter: VCM-D1; Uniblitz

Objective: 60X/1.0; Nikon

Objective: 10X/0.12; Zeiss

Home-made adaptor

Optical fiber: 500-2100 nm; F-MFC-C-1FC; Newport

Collimator: F-H10-VIS-APC; Newport

Software: Neuroplex 10.2; RedShirtImaging

Filters:  Dichroic:  532  nm;  Z532RDC  (for  laser).  Long  pass:  610  nm;  Q610LP  (for

voltage-sensitive dye, JPW3028); Chroma

Rectangular field stop: U-RFSS; Olympus

Camera port tube: C-mount; 0.25X; U-TV0.25XC; Olympus 

Multi-band glass dongle: 532 nm; YAMAMOTO

Precision spatial filter: ES01A; Onset

Post and holder: EP03N; 75 mm; Onset

Platform carrier: ECR01S; 77X42 mm; Onset

Optical bench: EOB01X; 25 cm; Onset

Base Clamps: EBC05S; Onset

 

(37) Transgenic mice

Strain
name: (1) B6.129-Asic1tm1Ccli
                     
(2) B6;129P2-Pvalbtm1(cre)Arbr/J
                     
(3) B6;129S5-Kltm1Lex/Mmucd
                     
(4) B6.Cg-Gt(ROSA)26Sortm14(CAG-tdTomato)Hze/J
                     
(5) B6;FVB-lfi208 Tg(Cspg4-cre)1Akik/J
                     
(6) B6N.129-Rpl22 tm1.1Psam/J
                     
(7) C57BL/6-Tg(Grik4-cre)G32-4Stl/J
                     
(8) Cnr1tm1Zim
                     
(9) ICR.Cg-Gad1<tm1.1Tama>
                     
(10) STOCK Ccktm1.1(cre)Zjh/J
                     
(11) STOCK Gad2 tm2(cre)Zjh/J
                     
(12) STOCK Ssttm2.1(cre)Zjh/J
                     
(13) STOCK Tg(Cspg4-DsRed.T1)1Akik/J
                     
(14) Tg(Calcrl,cre)4688Nkza
                     
(15) Viptm1(cre)Zjh/J
                     
(16) STOCK Tg(mI56i-flpe)39Fsh/J
                     
(17) B6(Cg)-Crhtm1(cre)Zjh/J
                      (18) STOCK Slc17a6tm2(cre)Lowl/J
                      (19) STOCK Tg(Prkcd-glc-1/CFP,-cre)EH124Gsat/Mmucd
                      (20) STOCK Fos tm2.1(icre/ERT2)Luo/J
                     
(21) STOCK Gli1 tm3(cre/ERT2)Alj/J
                     
(22) B6J.129S6(FVB)-Slc32a1tm2(cre)Lowl/MwarJ
                     
(23) Wnk1tm1Clhu
                     
(24) B6.Cg-Tg(Fos-tTA,Fos-EGFP*)1Mmay/J
                     
(25) B6.FVB(Cg)-Tg(Drd2-cre)ER44Gsat/Mmucd
                     
(26) B6.129-Grm5tm1.1Jixu/J

 


(38)  Software [Details]

                   Name: (1) Coreldraw- X3/X8 (10 quotas) [X8 free-trials]

                               (2) Ethovision- 3.1 (Owned by Dr. Irene ‘s lab)

                               (3) ImageJ (Fiji)- 1.47 and 1.5b [Download 1] [Download 2]

                               (4) LAS AF Lite- 4.0 [Download]

                               (5) Metamorph- NX

                               (6) Minianalysis- V 6.0.9

                               (7) NeuroExplorer- V 5 [Download]

                               (8) Neurolucida Explorer- V 10

                               (9) Neuromantic- V 1-6-3 [Download]

                               (10) Neuron- V 7.4 [Download]

                               (11) pClamp- V 10.2-10.6

                               (12) Prism- V 5 and 6

                               (13) Spike 2- V 5.08

                               (14) MetaFlour Guide  [Download]

 

(39)  Neuronal tracers [Details]

 

(40)  Air compressor

        All air compressors in Lien lab are "SIL AIR, Super Silent 30-TC" . [Link 1] [Link 2]

 

(41)  Trouble-shooting

        Stuck of the LN manipulator [Link]

                         

 

實驗室小鼠報表: https://goo.gl/hviJkI

 

訂貨及記錄查詢: https://goo.gl/t6GTdJ

 

化學藥品清單: [Link]