Bacterial biofilms are one of the most important challenges that modern medicine faces due to the difficulties of diagnosis, antibiotic resistance, and protective mechanisms against aggressive environments. For these reasons, methods that ensure the inexpensive and rapid or real-time detection of biofilm formation on medical devices are needed. This study examines the possibilities of using optical- and fiber-based biosensors to detect and analyze early bacterial biofilms. In this study, the biofilm-forming model organism Pseudomonas aeruginosa was inoculated on the surface of the optical sensor and allowed to attach for 2 h. The biosensors were made by a fiber-tip ball resonator, fabricated through a CO₂ laser splicer on a single-mode fiber, forming a weak reflective spectrum. An optical backscatter reflectometer was used to measure the refractive index detected by the sensors during different growth periods. The early biofilm concentration was determined by crystal violet (CV) binding assay; however, such a concentration was lower than the detection limit of this assay. This work presents a new approach of biofilm sensing in the early attachment stage with a low limit of detection up to 10⁻⁴ RIU (refractive index units) or 35 ± 20 × 10³ CFU/mL (colony formed units).

Optical fiber ball resonators based on single-mode fibers in the infrared range are an emerging technology for refractive index sensing and biosensing. These devices are easy and rapid to fabricate using a CO₂ laser splicer and yield a very low finesse reflection spectrum with a quasi-random pattern. In addition, they can be functionalized for biosensing by using a thin-film sputtering method. A common problem of this type of device is that the spectral response is substantially unknown, and poorly correlated with the size and shape of the spherical device. In this work, we propose a detection method based on Karhunen−Loeve transform (KLT), applied to the undersampled spectrum measured by an optical backscatter reflectometer. We show that this method correctly detects the response of the ball resonator in any working condition, without prior knowledge of the sensor under interrogation. First, this method for refractive index sensing of a gold-coated resonator is applied, showing 1594 RIU⁻¹ sensitivity; then, this concept is extended to a biofunctionalized ball resonator, detecting CD44 cancer biomarker concentration with a picomolar-level limit of detection (19.7 pM) and high specificity (30–41%).