Introduction Pre-analytical errors in cytology laboratories can significantly impact the accuracy of diagnostic results and turnaround times, ultimately affecting patient care. This article presents an evaluation of pre-analytical errors and proposes fostering strategies to enhance accuracy and efficiency in the cytology laboratory of a tertiary care hospital. The background discusses the importance of pre-analytical processes in ensuring reliable cytological diagnoses and the common errors encountered in specimen collection, handling, and transportation. Strategies for error reduction and improvement in turnaround times include staff education, standardization of procedures, utilization of appropriate collection and transport devices, implementation of quality control measures, and utilization of automation technologies. By addressing pre-analytical errors and implementing fostering strategies, cytology laboratories can optimize diagnostic accuracy, improve patient care outcomes, and enhance overall laboratory efficiency. Aims and objectives This study aims to assess the prevalence and nature of pre-analytical errors in the cytology laboratory of a tertiary care hospital to understand the extent of the issue, identify the specific factors contributing to pre-analytical errors like specimen collection, handling, and transportation processes, and pinpoint areas for improvement. It seeks to evaluate the impact of pre-analytical errors on the accuracy of cytological results and the efficiency of turnaround times, highlighting the consequences for patient care. Furthermore, the study aims to develop targeted strategies to minimize pre-analytical errors and enhance the accuracy of cytological results. Materials and methods This study was conducted at the Cytology Laboratory of our hospital from January 2023 to December 2023 after getting proper approval from the Institutional Review Board (IRB approval number 101/02/2024/PG/SRB/SMCH). It is a retrospective analytical study, and a total of 5412 samples from patients of the outpatient (OP) department, inpatient (IP) department, and community health outreach program facilities received in the cytology laboratory were analyzed during the study period. The inclusion criteria were the test samples sent specifically for cytological analysis. The samples sent for biochemical or microbiological examination were excluded. The frequency of sample distribution and rejected samples were calculated and the results were correlated. Results A total of 5,412 samples received in the cytology laboratory were analyzed during the study period. The majority of the samples were Papanicolaou smears (2,352, 43.5%), followed by fluid cytology (1,008, 18.6%) and ultrasound-guided fine-needle aspiration cytology (FNAC, 984, 18.2%). Of the total number of samples, 225 (4.16%) were repeated and 27 (0.5%) were rejected. Conclusions Pre-analytical, analytical, and post-analytical processes are the three key factors that determine the dependability and precision of cytological test results. Detecting critical alerts such as the positivity of malignancy underscores the paramount importance of result accuracy. Implementing good laboratory practices and conducting both external and internal audits can reduce the frequency of preventable errors in a cytology laboratory, thereby ensuring enhanced precision and expedited outputs.