世界各地的毒理学实验室都采用了眼表面刺激试验. With straightforward protocols and simple yet highly predictive endpoints, EpiOcular assays are routinely utilized throughout the cosmetic, personal care, chemical and pharmaceutical industries. EpiOcular是在20多年前开发的，已用于测试数千种材料. 科学家们是否需要一种风险评估方法来评估眼部传递的物质的刺激潜力，或者对世界各地运输的化学物质进行危害评估, EpiOcular is the test system of choice to meet your ocular testing needs.
Ocular Irritation Tests
EpiOcular Eye Irritation Test (OECD TG 492)
EpiOcular EIT是一种体外危害评估试验，能够区分非眼睛刺激物和眼睛刺激物. 企业所得税适用于广泛的有机和无机化学品，并被监管机构认可为 OECD TG 492.
EpiOcular Eye Irritation Test Protocol
观看如何进行表眼刺激试验(EIT - OECD TG 492)的视频:
EpiOcular Sub Draize Ultra-Mildness Test
眼表超轻度试验是一种体外风险评估试验，常规用于轻度材料，而Draize试验对轻度材料不敏感. This highly reproducible assay allows for quantifiable discrimination among mild, milder, and mildest product formulations.
EpiOcular MTT ET-50 (Neat)
这种体外风险评估测定是基于材料(直接应用)导致组织活力降低50% (ET-50)的有效时间。. Based on the ET-50, 测试物品被分为4个类别，从无刺激性到严重/极端，对应于兔子视力评分(MMAS)的分组。.
EpiOcular MTT ET-50 Neat Protocol
EpiOcular MTT ET-50 (Dilution)
Similar to the MTT ET-50 Neat method, 这种体外风险评估试验是基于物质(在水中稀释到20%)导致组织活力降低50% (ET-50)的有效时间。. Based on the ET-50, 测试物品被分为4个类别，从无刺激性到严重/极端，对应于兔子视力评分(MMAS)的分组。. This method is recommended for surfactant-based solutions and is applicable to water-soluble materials with a specific gravity of > 0.95.
EpiOcular MTT ET-50 Dilution Protocol
Browse our reference library to see how our EpiOcular tissue has been used in these areas of study.
Tissue:
Kit: 标准EpiOcular™试剂盒(OCL-200)由24个组织组成，每个组织直径9毫米. (Tissue “kits” contain tissues, a small amount of culture medium, and plasticware; contact 正规的网博软件有哪些 for specific kit contents.)
Substrate: Non-coated, 9 mm ID single well tissue culture plate inserts are used (e.g. Millicell PCF, Nunc polycarbonate single well tissue culture plate inserts).
Culture: At air liquid interface.
Histology: 5-8 cell layers (basal layer to apical surface).
Cornification: None.
Lot Numbers: 每个技术员每周生产的纸巾批号被分配到一个特定的批号. 通常，任何一周的纸巾生产都有多个批号. 在批号的末尾附加一个字母，以区分给定批号中的单个试剂盒. All tissue kits within a lot are identical in regards to cells, medium, handling, culture conditions, etc.
Shipment: At 4°C on medium-supplemented, agarose gels in 24-well plate.
Shipment day: Every Monday. Shipment on Thursday also possible upon special request.
Delivery: Tuesday morning via FedEx priority service (US). Outside US: Tuesday-Thursday depending on location.
Shelf life包括运输时间在内，纸巾在使用前可在4°C下保存4天. 但是，除非绝对必要，否则不建议延长储存期. In addition, 如果组织在同一天连续使用，将获得最佳的再现性, e.g. Tuesday afternoon or following overnight storage at 4°C (Wednesday morning).
Alternative tissues:
OCL-200-ABF: Antibiotic-free tissue. For last 3 days of culture, gentamicin is omitted from culture medium. Customer receives 200 ml of OCL-200-ASY-AFAB medium.
OCL-200-AFF: Anti-fungal-free tissue. For last 3 days of culture, Amphotericin is omitted from culture medium. Customer receives 200 ml of OCL-200-ASY-AFAB medium.
OCL-200-AFAB: Antibiotic, anti-fungal-free tissue. 培养的最后3天，培养基中不使用庆大霉素和两性霉素. Customer receives 200 ml of OCL-200-ASY-AFAB medium.
OCL-200-EACH: Single EpiOcular tissue insert.
OCL-200-FRZN-EA: Single OCL-200 tissue that has been subjected to 3 freeze/thaw cycles. Designed to serve as control for MTT auto-reduction test.
OCL-212: Half kit (12 cultures instead of standard 24).
OCL-606: Same as OCL-200 except 6 tissues per kit, each tissue cultured in a larger cell culture insert (22 mm diameter).
Cells:
Type: Normal human epidermal keratinocytes (NHEK).
Genetic make-up: Single donor.
Derived from: Neonatal-foreskin tissue.
Screened for: HIV, Hepatitis-B, Hepatitis-C, mycoplasma.
Medium:
Base medium: Dulbecco’s Modified Eagle’s Medium (DMEM).
Growth factors/hormones: Epidermal growth factor, insulin, 氢化可的松和其他非角化上皮分化的专有刺激物.
Serum: None.
Antibiotics: Gentamicin 5 µg/ml (10% of normal gentamicin level).
Anti-fungal agent: Amphotericin B 0.25 µg/ml.
pH Indicator: Phenol red.
Alternatives: Phenol red-free, antibiotic-free, anti-fungal-free, or hydrocortisone-free medium and tissue are available. Agents are removed at least 3 days prior to shipment.
Maintenance medium: Most assays with EpiOcular™ are performed within 24 hours. However, for longer experiments, 组织可以保存在OCL-200-MM(与EpiOcular™检测培养基相同)中。.
Quality Control and Sterility:
Visual inspection: All tissues are visually inspected and if physical imperfections are noted, tissues are rejected for shipment.
End-use testing: Tissues are exposed to 0.3% Triton X-100 for 5, 20, and 60 minutes. 使用MTT法确定每批组织降低组织活力(ET-50)所需的暴露时间(见正规的网博软件有哪些 EpiOcularTM ET-50方案). ET-50’s must fall within the range of the 1996 EpiOcular™ database: 12.2-37.5 minutes. ET-50’s in customers’ labs are often very similar to 正规的网博软件有哪些 results.
Sterility: All media used throughout the production process is checked for sterility. 维持培养基加或不加抗生素孵育1周，检查无菌性. 用于运输的24孔板上的琼脂糖凝胶也孵育1周，并检查任何污染迹象.
Screening for pathogens: 所有细胞都经过筛选，使用PCR检测HIV、乙肝和丙肝呈阴性. 然而，没有任何已知的测试方法可以完全保证细胞是无病原体的. Thus, 这些产品和所有人类衍生产品应按照CDC-NIH手册中建议的BSL-2(生物安全等级2)或更高级别处理, “Biosafety in microbiological and biomedical laboratories,” 1998. 如需进一步帮助，请联系您的现场安全专员或正规的网博软件有哪些技术服务.
Notification of lot failure: If a tissue lot fails our QC or sterility testing, the customer will be notified and the tissues will be replaced without charge. Because our QC and sterility testing is done post-shipment, notification will be made as soon as possible (Under normal circumstances, ET-50 failures will be notified by Wednesday 5pm; sterility failures will be notified within 8 days of shipment).
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