孢子虫无明显运动细胞器，全部发育阶段均营寄生生活。生活史较复杂，有无性的裂体增殖、孢子增殖和有性的配子生殖两种生殖方式，并可以在同一个宿主或分别在两个不同宿主体内完成。寄生于人体的孢子虫主要有疟原虫、刚地弓形虫、卡氏肺孢子虫、隐孢子虫等。
疟原虫是营寄生生活的孢子虫，主要寄生于人及多种哺乳动物，少数寄生于鸟类和爬行类动物，目前已知有130余种。寄生于人体的疟原虫主要有间日疟原虫、恶性疟原虫、三日疟原虫和卵形疟原虫等。疟原虫的基本结构包括胞核、胞质和胞膜，用瑞氏或吉姆萨染色后，胞核呈紫红色，胞质为蓝色。疟原虫在人体内先后在肝细胞和红细胞内发育，而且其致病是在红细胞内期裂体增殖时发生的。疟原虫在红细胞内发育包括小滋养体、大滋养体、裂殖体和配子体四个阶段，可通过瑞氏或吉姆萨染色观察其不同时期的不同形态。
疟原虫的病原学检查主要分为厚、薄血膜染色镜检法和血沉棕黄层定量分析法(QBC)。厚血膜中原虫比较集中，易检获，但制片中红细胞容易溶解，原虫形态有所改变，虫种鉴别比较困难;薄血膜中红细胞不易被破坏，疟原虫形态结构较完整、典型，容易识别和鉴别虫种，但原虫密度低时容易漏检;故常采用一张玻片同时制作厚、薄两种血膜，用瑞氏、吉姆萨或吖啶橙染色查找疟原虫的存在。QBC分析法的原理是由于感染疟原虫的红细胞比正常红细胞轻，而比白细胞略重，离心分层后，集中分布于正常红细胞层的上部，白细胞层的下部，加入吖啶橙试剂后，用荧光显微镜观察结果，敏感度比普通镜检法高很多，更简便、快捷，但费用较高。
阿拉丁疟原虫染色液 (吖啶橙染色液) 由吖啶橙和磷酸盐等组成，主要是采用荧光染料吖啶橙进行厚薄血膜的染色，通过渗透、吸附和化合等作用与寄生虫或组织细胞的不同成分结合，成为带荧光的标本，再经过荧光光源的激发，使结合的荧光素的标本发出荧光，通过荧光显微镜在暗室中观察疟原虫的存在，本法简单、快捷，检出率高于常规法。该试剂仅用于科研领域。不适用于临床诊断或其他用途。
产品组分及储存条件：
M1510409
| Component
| 100mL
| Storage
|
M1510409A
| 吖啶橙贮存液
| 1mL
| 4℃. Store in the dark.
|
M1510409B
| 磷酸盐缓冲液
| 100mL
| RT |
自备材料：
1、蜡笔、甲醇、去离子水
2、载玻片、荧光显微镜、暗室、抗荧光淬灭封片剂
操作步骤(仅供参考)：
1、制作血膜：厚血膜比吉姆萨染色的厚血膜略薄，可用3μl血涂成直径为1.2~1.5cm的亚厚血膜；薄血膜则不宜过薄，可在涂片时推片以45°角涂成较厚的薄血膜。
2、配制吖啶橙染色液：0.1ml 吖啶橙贮存液加入9.9ml 磷酸盐缓冲液混合即成，可于4℃保存10~15天。
3、染色前，先用蜡笔在厚薄血膜间划一条横线。
4、用0.5~1ml甲醇固定薄血膜，清水溶去厚血膜上的血红蛋白。
5、吸取吖啶橙染色液滴加到血膜上，室温避光染色30s~120s，加盖玻片。
6、荧光显微镜 (激发滤光片波长488nm，阻断滤光片波长515nm) 暗室中观察，计数并拍照，不能及时观察的可用抗荧光淬灭封片剂封片，可延缓荧光的淬灭。
染色结果：
疟原虫和白细胞的核呈现黄绿色荧光，胞浆呈橘红色荧光。
注意事项:
1、涂制血膜用的载玻片使用前需经铬酸洗液处理。
2、染色用的血膜厚薄应符合要求。
3、吖啶橙染色液一般建议临用前稀释，不宜存放过久。
4、滴加染色液应以完全覆盖血膜为准。
5、为了您的安全和健康，请穿实验服并戴一次性手套操作。
6、试剂开封后请尽快使用，以防影响后续实验效果。
Sporozoans lack distinct locomotor organelles and are parasitic throughout all developmental stages. Their life cycle is relatively complex, involving two reproductive modes: asexual schizogony and sporogony, as well as sexual gametogony. These processes can be completed either in a single host or separately in two different hosts. Sporozoans parasitic in humans mainly include Plasmodium, Toxoplasma gondii, Pneumocystis jirovecii, Cryptosporidium and so on.
Plasmodium is a parasitic sporozoan that primarily infects humans and various mammals, with a few species parasitizing birds and reptiles. To date, more than 130 species have been identified. The main species parasitic in humans include Plasmodium vivax, Plasmodium falciparum, Plasmodium malariae and Plasmodium ovale. The basic structure of Plasmodium consists of a nucleus, cytoplasm and cell membrane. After staining with Wright's or Giemsa's stain, the nucleus appears purplish red and the cytoplasm appears blue. Plasmodium develops successively in liver cells and red blood cells within the human body, and its pathogenicity occurs during the erythrocytic schizogony stage. The development of Plasmodium in red blood cells includes four stages: ring-form trophozoite, mature trophozoite, schizont and gametocyte. The distinct morphological characteristics of each stage can be observed after staining with Wright's or Giemsa's stain.
The etiological examination of Plasmodium is mainly divided into two methods: thick and thin blood film staining microscopy, and quantitative buffy coat (QBC) analysis. In thick blood films, the parasites are relatively concentrated, making them easy to detect. However, red blood cells tend to lyse during sample preparation, resulting in morphological changes of the parasites and thus making species identification difficult. In thin blood films, red blood cells are not easily damaged, and the parasites maintain intact and typical morphological structures, which facilitates species identification. Nevertheless, light infections are prone to missed detection due to the low density of parasites. Therefore, it is common practice to prepare both thick and thin blood films on a single slide, followed by staining with Wright's stain, Giemsa's stain or acridine orange to detect the presence of Plasmodium. The principle of QBC analysis is that red blood cells infected with Plasmodium are lighter than normal red blood cells but slightly heavier than white blood cells. After centrifugation and stratification, the infected red blood cells are concentrated in the upper part of the normal red blood cell layer and the lower part of the white blood cell layer. After adding acridine orange reagent, the results are observed under a fluorescence microscope. This method has much higher sensitivity than conventional microscopy, and is more simple and rapid, but it is relatively costly.
Aladdin Plasmodium Staining Solution (Acridine Orange Staining Solution) is mainly composed of acridine orange and phosphate. It is primarily used for staining thick and thin blood films with the fluorescent dye acridine orange. Through the effects of permeation, adsorption and combination, the dye binds to different components of parasites or tissue cells to form fluorescent specimens. Excited by a fluorescent light source, the specimens bound with fluorochrome emit fluorescence. The presence of Plasmodium can then be observed under a fluorescence microscope in a dark room. This method is simple and rapid, with a higher detection rate than conventional methods. This reagent is for research use only and is not suitable for clinical diagnosis or other purposes.
Product Components and Storage Conditions:
M1510409
| Component
| 100mL
| Storage
|
M1510409A
| Acridine Orange Stock Solution
| 1mL
| 4℃. Store in the dark.
|
M1510409B
| Phosphate Buffer Solution
| 100mL
| RT |
Self-prepared Materials:
1. Wax pencil, methanol, deionized water
2. Glass slides, fluorescence microscope, darkroom, anti-fluorescence quenching mounting medium
Operating Procedures (For Reference Only):
1. Prepare Blood Films: The thick blood film should be slightly thinner than that used for Giemsa staining; 3 μl of blood can be spread to form a sub-thick blood film with a diameter of 1.2–1.5 cm. The thin blood film should not be overly thin; it can be prepared by spreading the blood at a 45° angle with a spreader to form a relatively thick thin blood film.
2. Prepare Acridine Orange Staining Solution: Add 0.1 ml of acridine orange stock solution to 9.9 ml of phosphate buffer solution and mix well. The prepared staining solution can be stored at 4℃ for 10–15 days.
3. Before staining, draw a horizontal line between the thick and thin blood films with a wax pencil.
4. Fix the thin blood film with 0.5–1 ml of methanol, and rinse the thick blood film with clean water to dissolve the hemoglobin.
5. Pipette the acridine orange staining solution and drop it onto the blood films. Stain at room temperature in the dark for 30–120 seconds, then cover with a coverslip.
6. Observe the slides in a darkroom under a fluorescence microscope (excitation filter wavelength: 488 nm; barrier filter wavelength: 515 nm), then count and take photos. For samples that cannot be observed immediately, seal the slides with anti-fluorescence quenching mounting medium to delay fluorescence quenching.
Staining Results:
The nuclei of Plasmodium and white blood cells show yellow-green fluorescence, and the cytoplasm shows orange-red fluorescence.
Precautions:
1. Glass slides used for preparing blood films should be treated with chromic acid cleaning solution before use.
2. The thickness of the blood films for staining should meet the required specifications.
3. It is generally recommended to dilute the acridine orange stock solution immediately before use; the diluted staining solution should not be stored for an extended period.
4. The volume of the staining solution added should be sufficient to completely cover the blood films.
5. For your safety and health, please wear a lab coat and disposable gloves during operation.
6. Please use the reagent as soon as possible after opening to avoid affecting the results of subsequent experiments.
D2603001
危险品化学品经营许可证（带存储）