單分子定序在基因組學、轉錄組學、臨床診斷、藥物開發以及癌症篩檢等多個科學與醫學領域中具有重要意義。在現有方法中，螢光定序由於其高精確性與效率而被廣泛應用於DNA定序。傳統的螢光標記技術通常依賴電荷耦合元件（CCD）相機，在定序過程中捕捉多像素圖像。

本研究提出了一種基於單一像素分析的螢光標記檢測方法。此方法具有多項優勢，包括在低訊號雜訊比條件下提高定序準確性並降低計算資源。此外，相較於傳統技術所利用的多像素(二維)定序，我們能充分利用硬體資源，進而實現更高的通量。本研究集中於利用負二項式分布與我們修正負二項分布的方程式對單分子合成過程進行建模，並結合最大概似估計和Viterbi演算法，實現了更可靠的訊號檢測。

本篇研究基於螢光與生化反應而建立的模型，為模擬實驗過程提供了一個穩健的框架，也增進了第三代定序裡，對螢光發射與訊號接收之間關係的理解與準確度。

Single molecule sequencing plays a significant role in advancing various scientific and medical fields, including genomics, transcriptomics, clinical diagnostics, drug discovery, and cancer screening. Among the available approaches, fluorescence-based sequencing is widely utilized in DNA sequencing due to its precision and efficiency. Traditional fluorescence labeling techniques often rely on charge-coupled device (CCD) cameras to capture images of multiple pixels during sequencing.

In this study, we introduce a fluorescence labeling detection method based on single-pixel analysis. This approach offers distinct advantages, including improved accuracy and reduced resource requirements, particularly under low signal-to-noise ratio conditions. Moreover, it enables higher throughput compared to conventional techniques. The research focuses on modeling the single molecule synthesis process using negative binomial and modified negative binomial distributions. By integrating maximum likelihood estimation and the Viterbi algorithm, the method achieves enhanced signal detection reliability.

This fluorescence-based model provides a robust framework for simulating experimental processes, contributing to a better understanding of the relationship between fluorescence emission and signal reception.

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