Cyanines were and are still used in industry, and more recently in biotechnology (labeling, analysis). Cyanines have many uses as fluorescent dyes, particularly in biomedical imaging. Depending on the structure, they cover the spectrum from IR to UV.
Cyanine is the non-systematic name of a synthetic dye family belonging to polymethine group. The word cyanin is from the English word “cyan”, which conventionally means a shade of blue-green (close to “aqua”) and is derived from the Greek κυάνεος/κυανοῦς kyaneos/kyanous which means a somewhat different color: “dark blue”.
Cy7 is a near-IR fluor that is invisible to the naked eye (Excitation/emission maximum 750/776 ㎚). It is used in in vivo imaging applications, as well as the Cy7.5 dye.
Cyanines(Cy) replace advantageously conventional dyes such as Fluorescein(FITC) and rhodamines (TRITC, RRX), yielding brighter and more stable fluorescence.
시아닌은 여전히 업계에서, 그리고 최근에는 생명 공학 (라벨링, 분석)에서 사용되고 있습니다. 시아닌은 형광 염료로서, 특히 생체 의학 이미징에서 많은 용도로 사용되며 구조에 따라 IR에서 UV까지 스펙트럼을 커버합니다.
Cyanine은 polymethine 그룹에 속하는 합성 염료 계열의 비 계통적 이름입니다. cyanin이라는 단어는 일반적으로 청록색 ( “아쿠아”에 가까운)의 그늘을 의미하는 영어 단어 “cyan”에서 유래되었으며 다소 다른 색상 “진한 파란색 “을 의미하는 그리스어 κυάνεος / κυανοῦς kyaneos / kyanous에서 파생되었습니다.
Cy7은 육안으로는 보이지 않는 근적외선 형광체이며, Cy7.5 염료와 함께 생체 내 이미징 어플리케이션에 사용됩니다.
Cyanines (Cy)는 Fluorescein (FITC) 및 rhodamines (TRITC, RRX)와 같은 기존의 염료를 대체하여 보다 밝고 안정한 형광을 생성합니다.
Alexa Fluor 594 dye is a bright, red-fluorescent dye that can be excited using the 561 nm or 594 nm laser lines. For stable signal generation in imaging and flow cytometry, Alexa Fluor 594 dye is pH-insensitive over a wide molar range.
Cyan Fluorescent Protein (CFP) is a versatile biological marker for monitoring physiological processes, visualizing protein localization, and detecting transgenic expression in vivo. CFP can be excited by the 405 nm laser line and is optimally detected at 485 nm.
Cy5 dye is a bright, far-red-fluorescent dye that can be excited using the 633 or 647 nm laser lines, and visualized Cy5 filter sets. In addition to immunocytochemistry applications, Cy5 is also commonly used to label nucleic acids.
A variety of cyanine 5.5 (Cy5.5) dyes has been used to label biological molecules for fluorescence imaging and other fluorescence-based biochemical analysis. They are widely used for labeling peptides, proteins and oligos etc. Cy5.5 dyes are one type of the most common red fluorophores.
Cy7 is a near-IR fluor that is invisible to the naked eye (Excitation/emission maximum 750/776 nm). It is used in in vivo imaging applications, as well as the Cy7.5 dye.
DAPI (4′,6-diamidino-2-phenylindole) is a fluorescent stain that binds strongly to A-T rich regions in DNA. It is used extensively in fluorescence microscopy. As DAPI can pass through an intact cell membrane, it can be used to stain both live and fixed cells, though it passes through the membrane less efficiently in live cells and therefore the effectiveness of the stain is lower.
Fluorescein isothiocyanate (FITC) is a derivative of fluorescein used in wide-ranging applications including flow cytometry. FITC is the original fluoresce in molecule functionalized with an isothiocyanate reactive group (-N=C=S), replacing a hydrogen atom on the bottom ring of the structure. This derivative is reactive towards nucleophiles including amine and sulfhydryl groups on proteins.
The green fluorescent protein (GFP) is a protein composed of 238 amino acid residues (26.9 kDa) that exhibits bright green fluorescence when exposed to light in the blue to ultraviolet range. Although many other marine organisms have similar green fluorescent proteins, GFP traditionally refers to the protein first isolated from the jellyfish Aequorea victoria. The GFP from A. victoria has a major excitation peak at a wavelength of 395 nm and a minor one at 475 nm. Its emission peak is at 509 nm, which is in the lower green portion of the visible spectrum.
tdTomato is a genetic fusion of two copies of the dTomato gene (2) which was specifi cally designed for low aggregation (1). Its tandem dimer structure plays an important role in the exceptional brightness of tdTomato (Table I). Its excitation and emission maxima occur at 554 nm and 581 nm, respectively (1). Because tdTomato forms an intramolecular dimer, it behaves like a monomer, and has been used successfully for N- and C- terminal fusions. It shows excellent photostability and its half-time (t0.5) for maturation is one hour at 37°C.
Tetramethylrhodamine (TRITC) is a bright orange-fluorescent dye with excitation ideally suited to the 532 nm laser line. It is commonly conjugated to antibodies and proteins for cellular imaging applications.
Mercury : 형광 현미경법에 가장 흔히 사용하는 광원으로, 자외선 및 가시광선 영역에서 아주 밝다.
Spectrum of mercury
Xenon : 가시광선에서 상대적으로 연속적인 스펙트럼을 가진다. 염료 및/또는 시료의 스펙트럼 특성을 정량적으로 분석하는 시스템에서는 제논 램프를 사용하는 것이 좋지만, 같은 와트수의 수은 램프만큼 밝지는 않다.
Spectrum of Xenon
LED : 약 10 ~ 40nm 범위의 상대적으로 좁은 스펙트럼 대역폭 내에서 빛을 방출한다는 고유의 특징이 있다. 일반적으로 정격 수명이 10,000시간 이상이고 50,000시간 이상 사용하는 경우도 많고, 전체 정격 수명에 걸쳐 출력 강도가 비교적 일정하게 유지된다.
Spectrum of LED
Laser : 레이저
보통은 단색광의 좁은 스펙트럼 대역에서 극히 밝다. 하지만 레이저광의 간섭성으로 인해 샘플 영역에 균일한 조도로 비추기 어렵지만, Confocal 현미경에서는 갈바노 미러를 이용한 스캔 방식을 이용하여 이미지 획득시에 사용되어진다(LSCM). Spectrum of Laser
Metal-halide :
유효수명이 최대 2000시간으로, 일반적인 수은 램프의 200시간이나 제논 램프의 400시간과 비교하면 훨씬 길다.
AcquCAM Pro/S3 USB 3.0 camera is the perfect solution for many bio, industrial, medical applications. Utilizing the very sensitive mono CCD sensors, these cameras feature a specialization of fluorescence observations. With up to 30 fps and a trigger delay of less than 5 micro seconds.
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