染色原理 Principle
完整細胞膜可排除 Trypan Blue；膜完整性受損的細胞呈藍色。此方法量測的是染劑排除能力，結果受染色時間、細胞種類及顯微影像品質影響。Trypan Blue exclusion reflects membrane integrity. Dye uptake, cell type, staining time and imaging conditions affect classification.
CELL CULTURE · MICROSCOPY · EDUCATIONAL v0.1
Web-Trypan Blue
細胞存活率影像分析 Cell Viability Image Analyzer
上傳 Trypan Blue 染色的明視野顯微鏡照片，辨識候選細胞並依藍色染色特徵初步分類；逐顆人工覆核後計算存活率。支援選取 ROI、計數盤濃度換算，以及標註圖片與 CSV 匯出。Upload a brightfield Trypan Blue image, detect candidate cells, inspect staining-based classifications, correct each marker, and calculate cell viability. ROI selection, optional chamber concentration and local exports are included.
教學原型 · Educational prototype影像辨識採自行開發的傳統影像處理法，尚未經準確度驗證。染色、離焦、群聚與格線可能導致誤判；所有計數需人工確認。不得用於診斷、製程放行或臨床決策。
Rule-based computer vision; accuracy has not been validated. Focus, staining, overlap and grid lines may affect results. Manually verify all counts. Not for diagnosis or clinical decisions.
Rule-based computer vision; accuracy has not been validated. Focus, staining, overlap and grid lines may affect results. Manually verify all counts. Not for diagnosis or clinical decisions.
01 載入與設定 Upload & Settings
首次分析建議先用合成範例。深色染色細胞、背景亮度及相機白平衡都會影響判斷。Try the synthetic reference first. Results depend on dye appearance, illumination, and white balance.
02 人工覆核 Manual Review
ROI 模式拖曳矩形指定分析區；其餘模式點選既有標記可重新分類，點選空白處可新增標記。Drag to draw a rectangular ROI. In annotation modes, tap a marker to reclassify it or tap empty space to add a cell.
請上傳影像，或開啟合成示範。Upload an image or load the synthetic demo.
03 顯微影像 Microscopy Viewer
活細胞 / Live死細胞 / Dead待確認 / ReviewROI
04 統計與計算 Results & Calculations
活細胞 Live0
死細胞 Dead0
待確認 Review0
細胞存活率 Viability—
Cell viability (%) = Live ÷ (Live + Dead) × 100
待確認細胞不納入存活率分母；若存在待確認標記，數值為暫定結果。Unreviewed markers are excluded from the denominator. Results remain provisional until all markers are reviewed.
選用：計數盤濃度換算 Optional: Hemocytometer concentration
請先框選實際已知計數面積；完整 Neubauer 1 × 1 mm 大格，面積 1 mm²，標準深度 0.1 mm。若照片未校正計數區，請勿將結果作為細胞濃度。Select a physically known count area. A full Neubauer large square is 1 mm² at 0.1 mm depth. Uncalibrated fields cannot provide defensible cell concentrations.
活細胞濃度 Viable cells / mL—
總細胞濃度 Total cells / mL—
cells/mL = Count × dilution factor × 1000 ÷ (area [mm²] × depth [mm])
方法與限制 Method & Limitations
使用局部亮度閾值、形態學與連通區篩選尋找候選細胞，再以中心區域 B−R 色差推估藍色染色。相鄰細胞、離焦、染色沉澱及格線可能造成漏計或誤計；人工標記會保留在匯出檔。Local contrast, morphology and connected components detect candidates. Central blue chroma supports provisional classification. Overlaps, defocus, debris and grid lines remain sources of error.
文獻與程式說明 References & Source
Kuijpers L et al. PLoS ONE (2023), doi:10.1371/journal.pone.0291625；NIST Sources of error in cell viability measurement；Trypan Blue Exclusion protocol.
本站採原創前端實作，未複製上述研究的模型、標註資料或第三方 ImageJ 巨集。Original browser-only implementation. No third-party model, dataset or ImageJ macro is bundled.