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研究人員|中央研究院 細胞與個體生物學研究所

研究人員
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側邊選單開關 研究人員
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單分子生物核心實驗室

  • 黃婉媜Wan-Chen Huang
    研究助技師Assistant Research Scientist
    • 專長:Single-molecule biophysics Neuropharmacology Signal transduction
    • 信箱:wchuang1@gate.sinica.edu.tw
    • 電話:02-2787-1536
    • 位置:R526/ICOB
實驗室簡介展開收合
單分子生物核心實驗室(共2張照片)

The main purpose of establishing the Single-Molecule Biology Core (SMBC) laboratory is to provide a user-friendly platform that bridges the gap between biology and biophysics. The biophysical experiments that are coordinated by the SMBC may be applied to many of the various interdisciplinary areas within Academia Sinica. The service encompasses several functions, including protein purification, determining the physical chemistry properties of fluorescence molecules, single-cell or single-molecule imaging, and probing the interactions between single molecules. Facilities for the SMBC have been adapted from an academician, Dr. Tao-Shih Hsieh's laboratory and are described below. The SMBC aims to maintain all of the unique facilities that were established by Dr. Hsieh and to provide a continued source of information, expertise, and service, which will allow researchers with complete or limited knowledge of these techniques to apply the methods to their research.

服務申請流程及洽詢方式 設施使用規則說明 設備使用申請書 線上預約 收費標準及方式

設備介紹

單分子核心實驗室設備中屬貴重儀器者包括時間解析共軛焦螢光顯微鏡 (Time-Resolved Confocal Fluorescence Microscope) 及全內反射螢光顯微鏡 (Total Internal Reflection Fluorescence Microscopy);非貴重儀器則包括螢光分光光譜儀 (Spectrofluorometer) 及蛋白質純化系統 (Fast Performance Liquid Chromatography)。除包含細胞影像服務外,主要提供 DNA/RNA machinery的動態研究, 蛋白質折疊的動態與功能性研究以及單分子間或DNA/RNA和蛋白質間的交互作用機制探討..等等。另外還有完整的細胞培養室 (Cell Culture Room), 方便使用者進行細胞實驗,以減少因環境變化對細胞造成不必要的影響。

時間解析共軛焦螢光顯微鏡--Time-Resolved Confocal Fluorescence Microscope, Micro Time 200 (PicoQuant)

配備有三個常用波長(485、532、647 nm)的脈衝光源(pulsed laser),picosecond雷射控制 器,具備脈衝式交替激發功能(pulsed interleaved excitation, PIE),TCSPC & MCS積分系統,單光子偵測器(single photon counting detector)以及資料分析軟體。

時間解析共軛焦螢光顯微鏡

The MicroTime 200 is a time-resolved confocal microscope with single molecule sensitivity. Hence, numerous applications are possible with this instrument, including:

  • Single Molecule Spectroscopy/Detection
  • Time-Resolved Fluorescence
  • Fluorescence Lifetime Imaging (FLIM)
  • Fluorescence Correlation Spectroscopy (FCS)
  • Fluorescence Lifetime Correlation Spectroscopy (FLCS)
  • Fluorescence Resonance Energy Transfer (FRET)
全內反射螢光顯微鏡-- Total Internal Reflection Fluorescence Microscopy, CellTIRF (Olympus)

多光路全內反射顯微鏡包含三個常用波長(485、532、640 nm)的雷射光源,可自動調節入射光源角度及漸逝波穿透深度,電動載物台配備近紅外光可聚焦玻片表面,以利單分子定位; 並有EMCCD高敏感影像偵測系統,除提供細胞動態影像擷取外,更發展為適合單分子影像的偵測及動態分析。本實驗室建立此單分子系統並結合FRET的技術,主要用以偵測蛋白質與DNA分子間的交互作用:包括觀察解旋酶 (Helicase) 解開DNA雙股螺旋成單股的過程,以及抗癌藥物對拓樸異構酶 (Topoisomerase 2) 功能的動態干擾過程。

全內反射螢光顯微鏡

Total internal reflection fluorescence (TIRF) microscopy has become firmly established as a key technique in the investigation of molecular interactions at or near the cell surface. Hence, numerous applications are possible with this instrument, including:

  • Single Molecule Spectroscopy/Detection
  • Fluorescence Recovery After Photobleaching (FRAP)
  • Fluorescence Resonance Energy Transfer (FRET)
  • Cell Imaging
  • Calcium Imaging
螢光分光光譜儀 -- Spectrofluorometer,Fluorolog®-3 (Horiba JY)

量測螢光物理化學特性;包含吸收光及放射光光譜偵測,亦可利用內建偏光鏡片(polarizers),偵測分子之間的親和性差異以及利用螢光標識做為藥物篩選平台。

螢光分光光譜儀

多功能蛋白質純化及分析系統 -- Fast Performance Liquid Chromatography, AKTA purifier (GE Healthcare).

蛋白質純化及分析;為進行單分子間交互作用的研究,製備並純化大量重組蛋白質或螢光標識的蛋白質。

多功能蛋白質純化及分析系統

實驗室成員展開收合
姓名 職稱 電話 Email 備註
黃婉媜 研究助技師 27871536 wchuang1@gate.sinica.edu.tw
王堉璿 兼任研究助理 27871532 yswang1004@gmail.com
著作展開收合
1
Hsieh HW, Chen SC, Huang WC, Fang S, Yang EC, Hsu CC, Kou R. (2021) Social interactions upregulate hemolymph tryptophan and tyrosine levels in the male lobster cockroach. Horm Behav. 130:104935.
2
Muhammad Irham L, Chou WH, Wang YS, Adikusuma W, Sung-Ching Wong H, Aryani Perwitasari D, Huang WC, Chen BK, Yang HI, Chang WC. (2020) Evaluation for the Genetic Association between Store-Operated Calcium Influx Pathway (STIM1 and ORAI1) and Human Hepatocellular Carcinoma in Patients with Chronic Hepatitis B Infection. Biology (Basel). 9(11):388.
3
Irham LM, Wong HS, Chou WH, Adikusuma W, Mugiyanto E, Huang WC*, Chang WC. (2020) Integration of genetic variants and gene network for drug repurposing in colorectal cancer. Pharmacol Res. 161:105203.
4
Lee YC, Huang WC, Lin JH, Kao TJ, Lin HC, Lee KH, Lin HC, Shen CJ, Chang WC, Huang CC. (2018) Znf179 E3 ligase-mediated TDP-43 polyubiquitination is involved in TDP-43- ubiquitinated inclusions (UBI) (+)-related neurodegenerative pathology. J Biomed Sci.  25(1):76.
5
Huang WC*, Lee CY, Hsieh TS. (2017) Single-molecule FRET analysis discloses the dynamics of the DNA-topoisomerase II (TOP2) interaction in the presence of TOP2-targeting agents. Journal of Biological Chemistry.  292(30):12589-12598. (*Corresponding author)
6
Wei WC, Huang WC, Lin YP, Becker EBE, Ansorge O, Flockerzi V, Conti D, Cenacchi G, Glitsch MD. (2017) Functional expression of calcium-permeable canonical transient receptor potential 4-containing channels promotes migration of medulloblastoma cells. J Physiol595(16):5525-5544.
7
Lin S, Hong CH, Sheu BC, Wu LX, Huang WC, Huang WC, Guo CY (2016) Dynamic response of HPV16/anti-HPV16 pairs with unbinding events studied by atomic force microscopy. Biochemistry and Biophysics Reports. 6:203-208
8
Shen CJ, Chan SH, Lee CT, Huang WC, Tsai JP and Chen BK. (2016) Oleic acid-induced ANGPTL4 enhances head and neck squamous cell carcinoma anoikis resistance and metastasis via up-regulation of fibronectin. Cancer Letters386:110-122. (IF: 5.992)
9
Liao YH, Chiang KH , Shieh JM, Huang CR , Shen CJ, Huang WC*  and Chen BK*. (2016) Epidermal growth factor-induced ANGPTL4 enhances anoikis resistance and tumor metastasis in head and neck squamous cell carcinoma. Oncogene. DOI: 10.1038/onc.2016.371 (IF: 8.459, co-corresponding author)
10
Wong SC, Chang CM, Huang WC*, and Chang WC*. (2016) Genetic and molecular characteristics associated with local cytokines and related molecules across human cancers. OncoImmunology. 5(11):e1214789. (IF: 7.6, co-corresponding author)
11
Klahan S, Huang WC, Chang CM, Wong HS, Huang CC, Wu MS, Lin YC, Lu HF, Hou MF, Chang WC. (2016) Gene expression profiling combined with functional analysis identify integrin beta1 (ITGB1) as a potential prognosis biomarker in triple negative breast cancer. Pharmacol Res. 104:31-7. (IF:4.4)
12
Chang WC, Wu SL, Huang WC, Hsu JY, Chan SH, Wang JM, Tsai JP, Chen BK. (2015) PTX3 gene activation in EGF-induced head and neck cancer cell metastasis. Oncotarget.  6(10):7741-57. (IF:6.63)
13
Shieh JM, Shen CJ, Chang WC, Cheng HC, Chan YY, Huang WC, Chang WC, Chen BK. (2014) An increase in reactive oxygen species by deregulation of ARNT enhances chemotherapeutic drug-induced cancer cell death. PLoS One. 9(6):e99242. (IF:3.73)
14
Lin YS, Huang WC, Chen MS, Hsieh TS. (2014) Toward discovering new anti-cancer agents targeting topoisomerase IIα: a facile screening strategy adaptable to high throughput platform. PLoS One.  9(5):e97008. (IF:3.73)
15
Huang WC, Chai CY, Chen WC, Hou MF, Wang YS, Chiu YC, Lu SR, Chang WC, Juo SH, Wang JY, Chang WC. (2011)  Histamine regulates cyclooxygenase 2 gene activation through Orai1-mediated NFkB activation in lung cancer cells. Cell Calcium. 50:27-3. (IF:4.32)
16
Chou YH, Woon PY, Huang WC, Shiurba R, Tsai YT, Wang YS, Hsieh TJ, Chang WC, Chuang HY, Chang WC. (2011) Divalent lead cations induce cyclooxygenase-2 gene expression by epidermal growth factor receptor/nuclear factor-kappa B signaling in A431carcinoma cells. Toxicol Lett. 203(2):147-53
17
Huang WC, Chen ST, Chang WC, Chang KY, Chang WC, Chen BK. (2010) Involvement of aryl hydrocarbon receptor nuclear translocator in EGF-induced c-Jun/Sp1-mediated gene expression. Cellular and Molecular Life Science.  67(20):3523-33. (IF: 6.09)
18
Huang WC, Swietach P, Vaughan-Jones RD, Ansorge O, Glitsch MD. (2009) Differentiation impairs low pH-induced Ca2+ signaling and ERK phosphorylation in granule precursor tumour cells. Cell Calcium45(4):391-9. (IF:4.32)
19
Huang WC, Swietach P, Vaughan-Jones RD, Ansorge O, Glitsch MD. (2008) Extracellular acidification elicits spatially and temporally distinct Ca2+ signals. Current Biology.   18:781-5 (IF:10.988)
20
Su MC, Huang WC, Lien HC. (2008) Beta-catenin expression and mutation in adult and pediatric Wilm's tumors. APMIS.  116(9):771-8.
21
Huang WC, Young JS, Glitsch MD (2007) Changes in TRPC channel expression during postnatal development of cerebellar neurons. Cell Calcium42:1-10 (IF:4.32)
22
Wen HC, Huang WC, Ali A, Woodgett JR, Lin WW (2002) Negative regulation of phosphatidylinositol 3-kinase and Akt signalling pathway by PKC. Cellular Signalling. 15:37-45
23
Huang WC, Chio CC, Chi KH, Wu HM, Lin WW (2002)  Superoxide anion-dependent Raf/MEK/ERK activation by perxisome proliferator activated receptor gamma agonists 15-deoxy-delta(12,14)-prostaglandin J(2), ciglitazone, and GW1929. Exp Cell Res.  277(2):192-200.
24
Hsu YW, Chi KH, Huang WC, Lin WW (2001) Ceramide inhibits lipopolysaccharide-mediated nitric oxide synthase and cyclooxygenase-2 induction in macrophages: effects on protein kinases and transcription factors. The Journal of Immunology166(9):5388-5397